CA2274737A1 - Method and kit for hla class i typing - Google Patents

Method and kit for hla class i typing Download PDF

Info

Publication number
CA2274737A1
CA2274737A1 CA002274737A CA2274737A CA2274737A1 CA 2274737 A1 CA2274737 A1 CA 2274737A1 CA 002274737 A CA002274737 A CA 002274737A CA 2274737 A CA2274737 A CA 2274737A CA 2274737 A1 CA2274737 A1 CA 2274737A1
Authority
CA
Canada
Prior art keywords
seq
group
nucleic acid
hla
type
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
CA002274737A
Other languages
French (fr)
Inventor
Rainer H. Blasczyk
James Leushner
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Visible Genetics Inc
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of CA2274737A1 publication Critical patent/CA2274737A1/en
Abandoned legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Abstract

The present invention relates to methods and materials for determining the HLA
Class I type of a subject, wherein group-specific sequences are used to design primer molecules which may be used in amplification protocols which accurately identify the HLA group(s) and/or allele(s) carried by the subject.

Description

C ~ t~0.1_ Method And Kit For HL~. Class I Typing 1. it ductic n The present invention relates to rnethods and materials for determining the HLA Class I type of a subject, wherein group-specific sequences are used to design primer molecules which may be used in ,~rnplification protocols which accurately identify the HLA groups) and/or allc;le(s) carried by the subject.
2. Backg,~ound Of The Invention The Histocompatibility Locus Antigen ("HLA") Class I genes comprise three classical genes encoding the major transplantation antigens HLA-A, HLA-B, and HLA-C and seven other Class I genes of which HLA-E, HLA-F and HLA-G are probably functional genes and HLA-H, HLA-I, HLA-K and HLA-L are pseudogenes. The class I genes share a similar structure, which includes, inter alia, 5' -> 3', a 5' untranslated flanking region; a first e~:on ("exon 1 ") having a length of approximately 73 base pairs ("bp"); a first intron ("intron 1 ") having a length of approximately 130 bp; a second exon ("exon 2"), having a length of approximately 250 bp; a second intron ("intron 2"), having a length of approximately 272 bp;
a third exon ("exon 3"), having a length of approximately 276 bp; a third intron ("intron 3"), having a length of approximately 588 bp; and a fourth exon ("exon 4").
The HLA Class I genes are highly polymorphic among individuals. As of 1996, at least 73 alleles of HLA-A, 126 allelc;s of HLA-B and 35 alleles of HLA-C
have been identified. This variability is of particular relevance when tissue transplantation between a donor and a host is contemplated. The histocompatibility antigens of donor and host should be as similar as possible to avoid both immune rejection of the transplanted tissue as well as graft-versus-host disease. It is therefore important to accurately identify the HLA types of donor and host. In view of the exigencies implicit in tissue transplantation, it is desirable that the typing be accomplished as efficiently as possible.
Methods for determining alleles of HLA-A, HLA-Band HLA-C in a SUBSTITUTE SHEET ( rule 26 ) patient sample have been heavily investigated because of the functional importance of these genes in transplant tissue matching and autoimmune diseases. The first tests developed used immunological methods to identify epitopes expressed by various HLA loci. These tests (e.g., the complement-dependent cytotoxicity assay described in Terasaki and McClelland, Nature, 204:998, ( 1964)) identified broad serological specificities but were not capable of distinguishing between allelic members of a group, and sometimes mis-identified groups altogether. Unfortunately, even the most accurate of such low resolution assays cannot detect and distinguish all functionally significant transplant antigens (Anasetti et al. Hum. Immunol., 29:70 ( 1990)).
High resolution tests performed at the nucleic acid level which distinguish among alleles of each group have become the focus of recent research.
Current methods of high resolution typing include the following.
The Sequence Specific Oligonucleotide Probes ("SSOP") technique, as described in United States Patent No. 5,451,512 assigned to Hoffinan-La Roche, Inc., uses a reverse dot blot format, wherein HLA-A probes are immobilized on a membrane, and the labelled target (patient sample) DNA is hybridized to the membrane-bound probe (as described in Saiki et al., 1989, Proc. Natl. Acad.
Sci.
86:6230-6234). The pattern of hybridization to the probes on the dot-blot gives information regarding the HLA type of the individual. However, because hybridization is inherently not sufficiently specific to rule out minor differences in sequence between probe and patient sample, there is a possibility that the patient sample may contain an allelic variant which is not accounted for.
Another nucleic acid-based test is the Amplification Refractory Mutation System (ARMS) as described in the "HLA Class I SSP ARMS-PCR Typing Kit" Reference Manual, June 1995 edition, published by the Imperial Cancer Research Fund. This assay is based on the need for complementarity (matching) between the 3' end of an amplification primer and a target DNA sequence.
Absent such matching, the primer will not function properly and no fragment will be amplified. Sequence information is deduced by determining, for various pairs of primers acting on target DNA from a patient sample, whether or not a fragment is successfully amplified. The accuracy of the technique is limited by the number of SUBSTITUTE SHEET { rule 26 ) WO 98!26091 PCTICA9?/00955 primer pairs tested and by the possibility that allelic variations exist in regions of DNA which lie between the primers.
In order to overcome the foregoing shortcomings, it has been proposed that typing be accomplished by direct DNA sequencing (Santamaria et al., "HLA
Class I Sequence-Based Typing" Hum. Immunol. 37, 39-50 (1993); WO 9219771;
US Pat. 5,424,184). However, while direct sequencing of a patient's Class I
HLA
locus may conceptually be the most accurate, such sequencing may require a time-frame unsuitable for clinical practice. The success of direct sequencing methods may be expected to rely upon the design of efficient protocols and relevant primer sequences.
Prior to the present invention, diirect sequencing protocols have exhibited a number of disadvantages. For example, the method of Santamaria et al., supra, fails to provide sufficient information because it focuses on cDNA
(exon) sequences which, in view of exon sequence diversity, offer a very limited selection of conserved primer hybridization sites. In addition, because the Santamaria sequencing primers hybridize within an exon, they do not provide information for DNA
sequence upstream of the primer which is potentially decisive for distinguishing among alleles.
Further, the sites disclosed were determined before the recent discovery of dozens of more alleles that now need to be considered in identifying HLA type.
Intron sequences could provide; the preferred hybridization sites for amplification and sequencing primers for the :HLA-A, HLA-B and HLA-C genes because they may provide the DNA sequence of the full exon. Intron sequences for an HLA Class I gene were disclosed at least as early as 1985 (Weiss et al Immunobiol 170:367-380, (1985)). Due to their substantial diversity, and the difficulties in sequencing, few intron sequences have been published subsequently.
A number of researchers have made limited use of intron based oligonucleotides for limited aspects of HLA (Jlass I typing.
Blasczyk et al. (Tissue Antigens 1996: 47: 102-110) used exon based amplification primers to determine group specificity. After amplification, universal sequencing primers located in intron 2 were used to sequence the amplified fragment.
The paper does not disclose any intron sequence motifs from intron 1 or 3 or the 5' SUBSTITUTE SHEET ( rule 26 ) untranslated region.
Cereb et al. (Tissue Antigens 1995: 45:1-11), undertook the identification of intron sequences useful for locus-specific amplification primer sets for all Class I genes. These primer sets were designed to amplify all alleles of the same locus. No group specific amplification primers were sought or reported.
Further, amplified fragments were characterized by SSOP and not by direct sequencing.
Johnston-Dow et al (Poster Presentation: 1995 ASHI Meeting, Dallas, TX) presented a system for direct sequence determination of HLA-A wherein degenerate exon based primers were used to amplify exons 1 to 5 of the genomic HLA-A DNA sequence. As in Cereb et al., supra, the degenerate primer pool was designed to amplify all alleles of the HLA-A locus. Group specificity was not sought or reported. Further, sequencing of the amplified fragment was obtained using a degenerate primer mix wherein primers hybridize to intron regions flanking exons 2 and 3.
A rational approach to typing of classical HLA Class I loci would provide a simplified series of steps for high resolution typing of each allele of each loci in a patient sample using intron based oligonucleotides. Further, this method would be able to identify new alleles without ambiguities.
SUBSTITUTE SHEET ( rule 26 ) 3. Summary Of The Invention The present invention relates to materials and methods for high-resolution, nucleic acid-based typing of the three classical HLA Class I genes (comprising the loci HLA-A, HLA-B and HLA.-C) in a patient sample. It is based, in part, on the discovery of group-specific sequence motifs, derived from the analysis of numerous patient samples, which include sequences of the 5' flanking region, intron 1, intron 2, and intron 3. Such sequence motifs may be used to design amplification primers which may be used to identify the HLA group or type of a subj ect. The invention is also based, in part, on the determination of numerous allele-specific sequences which may be used to confirm the precise allelic type of a subject.
The present invention provides for substantially purified nucleic acids which are capable of selectively hybridizing with group specific sequence motifs in untranslated regions of the HLA-A, HLA-B or HLA-C gene loci. Such nucleic acids, which may be comprised in a kit, may be used, alone or in conjunction with exon-based primers, to determine the group specificity of HLA-A, HLA-B, or HLA-C
alleles contained in a patient sample and to idf;ntify the specific alleles present.
In particular embodiments, the present invention provides for methods of ascertaining the HLA Class I type of a subj ect which comprise performing a first amplification reaction which identifies the group type of the subject, and a second amplification reaction which produces allele-specific nucleic acids for sequencing.
3.1. Definitions "Allele" means one of the alternative forms of the gene in question;
"Amplification" means the process of increasing the relative abundance of one or more specific genes or gc;ne fragments in a reaction mixture with respect to the other genes. A method of amplification which is well known by those skilled in the art is the polymerase chain reaction (PCR) as described in United States Patents Nos. 4,683,194, 4,683,195 and 4,683,:202, which are incorporated herein by reference. The PCR process involves the use of pairs of primers, one for each complementary strand of the duplex DNA (wherein the coding strand is referred to as the "sense strand" and its complementary strand is referred to as the "antisense strand"), that will hybridize at a site located near a region of interest in a gene. Chain SUBSTITUTE SHEET ( rule 26 ) extension polymerization (without a chain terminating nucleotide) is then carned out in repetitive cycles to increase the number of copies of the region of interest many times. The amplified oligonucleotides are then separated from the reaction mixture and used as the starting sample for the sequencing reaction. Gelfand et al.
have described a thermostable enzyme, "Taq polymerase," derived from the organism Thermus aquaticus, which is useful in this amplification process (see United States Patent Nos. 5,352,600 and 5,079,352 which are incorporated herein by reference);
"Group" as used herein, refers to a subset of alleles of one loci, all of which share sequence features which distinguish them from other groups. For example, serological group reactivity {in a lymphocytotoxicity assay) is the conventional basis for nomenclature of HLA alleles. The first two digits of an allele refer to the serological group; for example, the designation A*0201, A*0202, A*0217 all are members of the A2 group. Further, typically the nomenclature refers to the serological split group (e.g., A23 and A24 are serological splits of A9;
"Group-specific sequence motif' means a generally short, 1-25 nucleotide ("nt") sequence of nucleic acid which is found only in one or a few groups.
Where a motif is shared by several groups in one region of the HLA locus, group-specific sequence motifs in other regions of the locus may serve as group-distinguishing features. The motif may share one or more nucleotides with the consensus sequence for the region;
"Haplotype" means the allele present on one chromosome;
"Heterozygote" means the presence of at least two different alleles of a gene;
"Homozygote" means the presence of a single species of allele of a gene;
"Locus" means a gene, such as HLA-A, HLA-B or HLA-C;
"Locus specific" means an event or thing associated with only one locus;
"Patient sample" means a sample collected from a patient in need of HLA typing which contains a sufficient amount and quality of nucleic acid (preferably DNA) for the performance of an amplification reaction. A
nonlimiting example of a suitable source is peripheral blood lymphocytes, tissue (including cell SUBSTITUTE SHEET ( rule 26 ) cultures derived therefrom, mucosal scrapes, spleen and bone marrow;
"Primer" means a polynucleotide generally of 5-50 nucleotides length which can serve to initiate a chain extension reaction;
"Sequencing" or "DNA sequencing" means the determination of the order of nucleotides in at least a part of a gene. A well known method of sequencing is the "chain termination" method first described by Sanger et al., Proc.
Nat'1 Acad.
Sci. (USA) 74(12): 5463-5467 (1977) (recently elaborated in EP-B1- 655506, and Sequenase 2.0 product literature (Amersham Life Sciences, Cleveland) incorporated herein by reference). Basically, in this process, DNA to be sequenced is isolated, rendered single stranded, and placed into four vessels. In each vessel are the necessary components to replicate the DNA strand, which include a template-dependant DNA polymerase, a short primer molecule complementary to a known region of the DNA to be sequenced, and individual nucleotide triphosphates in a buffer conducive to hybridization between the primer and the DNA to be sequenced and chain extension of the hybridized primer. In addition, each vessel contains a small quantity of one type of optionally detectably labeled dideoxynucleotide triphosphate, e.g., dideoxyadenosine triphosphate ("ddA"), dideoxyguanosine triphosphate ("ddG"), dideoxycytosine triphosphate ("ddC"), or dideoxythymidine triphosphate ("ddT"). In each vessel, each piece of the isolated DNA is hybridized with a primer. The primers are then extended, one base at a time to form a new nucleic acid polymer complementary to the isolated pieces of DNA. When a dideoxynucleotide is incorporated into the extending polymer, this terminates the polymer strand and prevents it from being furl:her extended. Accordingly, in each vessel, a set of extended polymers of specific lengths are formed which are indicative of the positions of the nucleotide corresponding to the dideoxynucleic acid in that vessel. These sets of polymers are then evaluated using gel electrophoresis to determine the sequence.
"Specific hybridization" mean:. hybridization of one strand of a nucleic acid to its complement.
"Target sequence" means the preferred site for specific hybridization of a primer; and SUBSTITUTE SHEET ( rule 26 ) "Untranslated region" refers to a portion of an HLA locus which is not transcribed into RNA and eventually translated into protein. Examples of untranslated regions are the 5' and 3' flanking regions and intron sequences. For example, the 5' flanking region is neither transcribed nor translated, and intron sequences are transcribed but not translated.
4. Description Of The Figures FIGURE 1 is an illustration of the principle for an HLA class I
sequencing strategy. Group-specific primers are used for PCR amplification, and universal primers located in the 2nd intron are used for sequencing, regardless of the amplified group. 5'FR= 5' flanking region; 5' UTR= 5' untranslated region (-1 to -23 from the ATG start codon in exon 1 ).
FIGURE 2A and 2B depict, in schematic form, a method of the invention in which a cocktail of HLA-A group specific primers is used to amplify target DNA contained in a patient sample. The products of amplification are then separated electrophoretically in an agarose gel, allowing the identification, by fragment mobility, of fragments corresponding to groups A2 and A3. Primers specific for groups A2 and A3 are then used to amplify duplicate samples of target DNA
in separate reactions, to produce A2 and A3 fragments which may then be sequenced using universal sequencing primers. FIGURE 2C and 2D depict a strategy wherein group type specificity is determined by reaction of aliquots of genomic DNA in separate reactions with a panel of primer pairs.
FIGURE 3 depicts the nucleic acid sequences of the HLA-A 5' flanking region in various alleles, including a consensus sequence (SEQ ID NO:
l ) as well as the sequences for the following alleles: A*0101 (SEQ ID N0:2); A*0301 (SEQ ID N0:3); A*1101 (SEQ ID N0:4); A*1102 (SEQ ID N0:5); A*3001 (SEQ ID
N0:6); A*3002 (SEQ ID N0:7); A*3004 (SEQ ID N0:8); A*0201-11 (SEQ >D
N0:9); A*0215 (SEQ LD NO:10); A*0217 (SEQ ID NO:11); A*6801 (SEQ ID
N0:12); A*6802 (SEQ ID N0:13); A*6901 (SEQ ID N0:14); A*2301 (SEQ ID
N0:15); A*2402 (SEQ ID N0:16); A*2403 (SEQ ID N0:17); A*2404 (SEQ ID
N0:18); A*2405 (SEQ ID N0:19); A*2407 (SEQ ID N0:20); A*2501 (SEQ ID
N0:21); A*2601 (SEQ ID N0:22); A*3402 (SEQ ID N0:23); A*4301 (SEQ ID
SUBSTITUTE SHEET ( rule 26 ) N0:24); A*6601 ID N0:25); (SEQID N0:26); A*6603 (SEQ A*6602 (SEQ ID

N0:27); A*2901 ID N0:28); (SEQID N0:29); A*31012 (SEQ A*2902 (SEQ ID

N0:30); A*3201 ID N0:31); (SIEQID N0:32); A*3303 (SEQ A*3301 (SEQ ID

N0:33); A*7401 ID N0:34); (SEQID N0:36); A*7403 (SEQ A*7402 (SEQ ID

N0:37); and A*8001 (SEQ ID N0:38).

FIGURE 4 depicts the nucleic acid sequences of HLA-A intron 1 in various alleles, including a consensus sequence: (SEQ ID N0:39) as well as the sequences for the following alleles: A*0101 (SEQ ID N0:40}; A*0301 (SEQ ID
N0:41 ); A* 1101 (SEQ ID NO:42); A* 1102 (SEQ ID N0:43); A*3001 (SEQ ID
N0:44); A*3002 (SEQ ID NO:45); A*3004 {SEQ ID N0:46); A*0201 (SEQ ID
N0:47); A*0202 (SEQ ID N0:44); A*0203 (SEQ ID N0:49); A*0204 (SEQ ID
NO:50); a*0205 (SEQ ID NO:51); A*0206 (SEQ ID N0:52); A*0207 (SEQ ID
N0:53); A*0207 {SEQ ID NO:54); A*0208 (SEQ ID NO:55); A*0209 (SEQ ID
N0:56); A*0210 (SEQ ID N0:57); A*0211 (S~EQ ID N0:58); A*0215 (SEQ ID
N0:59}; A*0217 (SEQ ID N0:60); A*6801 (S~EQ ID N0:61); A*6802 (SEQ ID
N0:62); A*6901 (SEQ ID N0:63); A*2301 (S~EQ ID N0:64); A*2402 (SEQ ID
N0:65); A*2403 (SEQ ID N0:66); A*2404 (SEQ ID NO:67); A*2405 (SEQ ID
N0:68); a*2407 (SEQ ID N0:69); A*2501 (S:EQ ID N0:70); A*2601 (SEQ ID
N0:71 ); A*3402 (SEQ ID N0:72); A*6601 (SEQ ID N0:73); A*6602 (SEQ ID
N0:74) A*6603 (SEQ ID N0:75); A*4301 (SEQ ID N0:76); A*2901 (SEQ ID
N0:77); A*2902 (SEQ ID N0:78); A*3101 (SEQ ID N0:79); A*3201 (SEQ ID
N0:80); A*3301 (SEQ ID N0:81); A*3303 (SEQ ID N0:82); A*7401 (SEQ ID
N0:83); A*7402 (SEQ ID N0:84); A*7403 (SEQ ID N0:85); and A*800I (SEQ ID
N0:86).
FIGURE 5 depicts the nucleic acid sequences of HLA-A intron 2 in various alleles, including a consensus sequence (SEQ ID N0:87) as well as sequences for the following alleles: A*0101 (SEQ ID NO:88); A*0201 (SEQ ID N0:89);
A*0202 (SEQ ID N0:90); A*0203 (SEQ ID rd0:91 ); A*0204 (SEQ ID N0:92);
A*0205 (SEQ ID N0:93); A*0206 (SEQ ID N0:94); A*0207 (SEQ ID N0:95);
A*0208 (SEQ ID N0:96); A*0209 (SEQ ID N0:97); A*0210 (SEQ ID N0:98);
A*0211 (SEQ ID N0:99); A*0215 (SEQ ID NO:100); A*0217 (SEQ ID NO:101);
SUBSTITUTE SHEET ( rule 26 ) A*6801(SEQID N0:102); (SEQID N0:103); (SEQ ID N0:104);
A*6802 A*6901 A*2501(SEQID NO:105); (SEQID N0:106); (SEQ ID N0:107);
A*2601 A*4301 A*6601(SEQID N0:108); (SEQID N0:109); (SEQ ID NO:l A*6602 A*6603 10);

A*3402(SEQID NO:111); (SEQID NO:112); (SEQ ID N0:113);
A*2901 A*2902 A*310I(SEQID N0:114); (SEQID NO:115); (SEQ ID N0:116);
A*3201 A*3301 A*3303(SEQID N0:117); (SEQID N0:118); (SEQ ID N0:119);
A*7401 A*7402 A*7403(SEQID N0:120); (SEQID N0:121); (SEQ ID N0:122);
A*2301 A*2402 A*2403(SEQID N0:123); (SEQID N0:124); (SEQ ID N0:125);
A*2404 A*2405 A*2407(SEQID N0:126); (SEQID N0:127); (SEQ ID NO:I28);
A*0301 A* 1101 A*1102(SEQID N0:129); (SEQID N0:130); (SEQ ID N0:131);
A*3001 A*3002 A*3004(SEQID N0:132);
and A*8001 (SEQ ID N0:133).

FIGURE 6 depicts the nucleic acid sequences of HLA-A intron 3 in various alleles, including a consensus sequence (SEQ ID N0:134) as well as sequences for the following alleles:
A*0101 (SEQ
ID N0:135);
A*0301 (SEQ
ID

N0:136); A* (SEQID N0:137); A* 1102 (SEQ ID N0:138);
1101 A*3001 (SEQ ID

N0:139); A*3002(SEQID N0:140); A*3004 (SEQ ID N0:141); A*0201 (SEQ ID

N0:142); A*0202(SEQID N0:143); A*0203 (SEQ ID N0:144); A*0204 (SEQ ID

N0:145); A*0205(SEQID N0:146); A*0206 (SEQ ID N0:147); A*0207 (SEQ ID

N0:148); A*0208(SEQID N0:149); A*0209 (SEQ ID NO:150); A*0210 (SEQ ID

NO:151); A*0211(SEQID N0:152); A*0215 (SEQ ID N0:153); A*0217 (SEQ ID

N0:154); A*6801(SEQID NO:155); A*6802 (SEQ ID N0:156}; A*6901 (SEQ ID

N0:157); A*2301(SEQID N0:158); A*2402 (SEQ ID N0:159); A*2403 (SEQ ID

N0:160); A*2404(SEQID N0:161 ); A*2405 (SEQ ID N0:162);
A*2407 (SEQ ID

N0:163); A*2501(SEQID N0:164); A*2601 (SEQ ID N0:165); A*3402 (SEQ ID

N0:166); A*4301(SEQID N0:167); A*6601 (SEQ ID N0:168}; A*6602 (SEQ ID

N0:169); A*6603(SEQID N0:170); A*2901 (SEQ ID N0:171); A*2902 (SEQ ID

N0:172); A*3101(SEQID N0:173); A*3201 (SEQ ID N0:174); A*3301 (SEQ ID

N0:175); A*3303(SEQID N0:176); A*7401 (SEQ ID N0:177}; A*7402 (SEQ ID

N0:178); A*7403(SEQID N0:179); and A*8001 {SEQ ID N0:180).

FIGURE 7 depicts a phyiogenetic tree of the 5' flanking and 5' untranslated regions of HLA-A.
SUBSTITUTE SHEET ( rule 26 ) FIGURE 8 depicts a phylogenetic tree of introns 1-3 of the HLA-A
gene.
FIGURE 9 depicts a phylogenetic tree of introns 1-3 of the HLA-B
gene.
FIGURE 10 depicts the results of amplification using group-specific exon region primers to determine HLA-A group type, wherein the group specificity is determined to be 6601 and 3201 (see Table 7).
FIGURE 11 depicts the results of amplification using group-specific exon region primers to determine HLA-A group type, wherein the group specificity is determined to be 020x and 680x (see Table 8).
5. Detailed Description Of The Invention The present invention relates to compositions and methods which may be used to efficiently and accurately determine the HLA Class I type of a patient sample.
The present invention is based.. in part, on the determination of group-specific sequence motifs in regions of HLA Class I loci. These motifs may be used to design oligonucleotides which may be used as group-specific primers in nucleic acid amplification reactions. The present invention is also based, in part, on the determination of the sequences of regions of a wide variety of alleles of HLA
Class I
loci; such sequences may be used to distinguish one allele from another.
Sequences of regions including the 5' flanking region of HI,A-A and introns l, 2 and 3 of HLA-A
are provided herein, and are set forth in Figuxes 3-6.
In general, the methods of the invention may be described as follows.
Comparison of nucleotide sequences of an HLA locus among members of an HLA
Class I group, which lie in either untranslated~ or exon regions, may be used to identify group-specific motif sequences. Identification of groups may be by establishing serological relationships or using phylogenet:ic information, as set forth in Figures 7-9. Based on the group-specific motif sequences, oligonucleotide primers may be designed, synthesized, and used to amplify a portion of the HLA locus.
Oligonucleotides used in this manner are referred to herein as "group-specific SUBSTITUTE SHEET ( rule 26 ) primers" and, in particular, as "group-specific untranslated region primers"
or "group-specific exon region primers", as the case may be.
In preferred nonlimiting embodiments of the invention, the primers correspond to untranslated regions of the HLA Class I locus ("group-specific untranslated region primers"). Such primers may be used in pairs, wherein each member of the pair hybridizes to an untranslated region lying on either side of at least one exon. For example, but not by way of limitation, primer pairs may be oligonucleotide pairs which hybridize to group-specific motifs in the 5' untranslated region and the first, second, or third intron; the first intron and the second or third intron; or the second and third intron.
The group-specific primers may be used in several different methods according to the invention. In a first series of nonlimiting embodiments, the group-specific primers may be used in a diagnostic manner to identify which allelic groups are present in a patient sample. In a second series of nonlimiting embodiments, the group-specific primers may be used to amplify sufficient amounts of a particular allelic fragment which is then subjected to direct nucleotide sequencing using universal sequencing primers.
According to the first series of embodiments, the present invention provides for a method of determining the HLA Class I group type of a subject comprising (i) combining a group-specific primer pair with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA
may occur; and (ii) determining whether a nucleic acid product is produced by the amplification; wherein the ability of a primer pair to produce a nucleic acid product is associated with a particular HLA group type. The group-specific primers may be group-specific exon region primers or group-specific untranslated region primers. In related embodiments the present invention provides for a method of determining the HLA Class I group type of a subject comprising (i) combining a plurality of group-specific exon region primer pairs with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur;
(ii) determining the size of the nucleic acid products of the amplification; and (iii) correlating the size of the product with the predicted size of a fragment associated SUBSTITUTE SHEET ( rule 26 ) with a particular HLA group type. The plurality of primers is referred to as an HLA
"cocktail" (see Figures 1 and 2). These first methods may be used to provide useful diagnostic information. For example, group typ~s determination may serve as a first level of comparison for a histocompatibility analysis, even without identification of the specific alleles) involved. For example, if av potential donor and host are being evaluated for tissue transplantation, if it is found that their group types do not match, no further comparison may be necessary. If, alternatively, their types do match, fur-ther analysis, for example by direct sequencing, may be desirable.
According to the second series of embodiments, the present invention provides for a method of determining the HLA Class I allelic type of a subject comprising (i) combining a group-specific oligonucleotide primer pair with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur; (ii) collecting the nucleic acid product of the amplification; and (iii) determining the nucleic acid sequence of the product.
The group-specific primer pair used may be determined based on the group type of the subject, as determined using the first method, described above. In preferred embodiments of the invention, group-specific untranslated region primers which span a region of the HLA locus containing allele-specific sequence may be utilized.
If a subject is heterozygous, separate amplification reactions are performed for each group identified (e.g., separate reactions to amplify fragment for group A2 and group A3;
see Figure 2). Sequencing may be performed using universal sequencing primers which will operate irrespective of HLA group or allelic type.
A more detailed description of the invention follows. Most alleles of the classical HLA Class I gene loci (consisting of HLA-A, HLA-B and HLA-C) can be distinguished on the basis of exon 2 and 3 alone. In one non-limiting embodi-ments, a method of the invention takes advantage of this fact, and employs the strategy generally described in Figure 2, using; the example of HLA-A. A
genomic DNA sample is prepared from a patient sample according to well known techniques.
Aliquots of the genomic DNA may then separately be reacted with a panel of group-specific exon region primer pairs (Figure 2C), wherein the successful amplification of a DNA fragment is associated with a particular group type. Alteratively, as depicted SUBSTITUTE SHEET ( rule 26 ) in Figure 2A), part of the sample may be treated with a cocktail of group-specific exon region primer pairs. Each primer pair in the cocktail will amplify only selected allelic groups because they specifically hybridize to group specific intron sequence motifs. Between them, under suitable polymerase chain reaction (PCR) conditions, the cocktail may amplify all known HLA-A groups, with each group specific amplification product having a different length. When reaction products are separated on an agarose gel the groups) present in the patient sample may be identified by length.
Optionally, once the group specificity is determined, the direct sequence of alleles may be determined for precise allelic identification. As illustrated in Figure 2 B), a further part of the patient sample DNA may be treated under PCR
conditions with a pair of primers that are specific for the previously determined group;
preferably such primers are group-specific untranslated region primers, which span greater distances of the locus. If two groups were detected, then two separate reactions are performed. At completion of the second amplification, the reaction products are sequenced using an intron based "universal primer" which hybridizes to an intron sequence which is conserved among all alleles of the locus. Though it is theoretically possible to use a sequencing primer which is specific for the amplified group only, it is found that using a universal primer simplifies the method and the preparation of a kit. Various universal sequencing primers are specifically provided herein (see infra) which hybridize, respectively, to intron sequences flanking the 5' end of exon 2, the 3' end of exon 2, the 5' end of exon 3 and the 3' end of exon 3.
The substantial advantage of the method of the invention is that the initial group specific amplification allows a PCR based separation of haplotypes in 95% of patient samples. The separation of the haplotypes is a major achievement of this protocol since it permits the resolution of cis/trans linkages of heterozygote sequencing results which cannot be achieved with other protocols. With the instant invention, a separation of the haplotypes may be achieved in serological heterozygous samples with the sequencing primer mixes ("PMs") described in Table 2 (infra) using group-specific amplification corresponding to the serological families. The selection of the PMs used for sequencing depends on the amplification patterns of the preceding SUBSTITUTE SHEET ( rule 26 ) PCR-SSP low-resolution typing. The primers a~-e designed to work with a in a single cycle protocol including, but not limited to, a PCR protocol on a Perkin Elmer System 9600, maintaining typing capacities of the laboratory. All PCR products carry sufficient sequence information for a complete subtyping. This approach is superior to a typing system using a single pair of generic primers followed by direct sequencing or SSO hybridization, even if the amplification strategy is locus-specific.
The substantial advantage of Sequence Based Typing (SBT) is the definition of the cis/trans linkage of sequence motifs. SBT after generic PCR amplification cannot define the cis/trans linkage of sequence motifs and therefore mimics oligotyping. The rapidly growing number of newly identified alleles confirms that new alleles have arisen mainly from gene conversion events which have usually taken place between different alleles of the same locus. Newly identified alleles are not characterized by new sequence motifs, but by a new combination of already existing sequence motifs.
From this observation it may be concluded that the amount of alleles at each locus may theoretically represent all possible combinations of known sequence motifs. Of course, some of them will fall victim to negative selection. Nevertheless, it can be expected that still an enormous amount of alleles are yet unidentified. PCR-SSP
subtyping strategies using a restricted number of oligonucieotides which do not cover all possible sequence motifs suffer from this limitation. If the cis/trans linkage of the analyzed polymorphic regions is not defined some new alleles may be mistyped as a heterozygous combination of known alleles. 'this has consequences with respect to SBT strategies. An unambiguous typing result of SBT after generic PCR
amplification is only unambiguous with regard to the presently known HLA
sequence databank. However, with the detection of new alleles this result can become ambiguous over the course of time. This observation has already been made in PCR
based DRB 1 typing during the last five years and will probably also occur in PCR
based class I typing. Considering the above points, the idea of the instant SBT
approach is not only to identify the HLA-A, HLA-B and HLA-C subtypes, but to cover as many of the polymorphic sites as possible and to define the ~is/trans linkage of the polymorphic sequence motifs. Typing results obtained with this method will remain unambiguous independently of the growing HLA sequence databank.
SUBSTITUTE SHEET ( rule 26 ) In general, group-specific primers are desirably designed to facilitate hybridization to their intended targets. It should be taken into account that homology between different groups, and indeed between group-specific motifs, may exist.
Accordingly, in preferred embodiments of the invention, a primer may be designed such that it hybridizes to its group target under relatively stringent conditions. For example, one or more mismatched residues may be engineered into the 3' domain of the molecule. Further, the primer may be designed such that it differs from any naturally occurring or consensus sequence, but rather has mismatches inserted which serve to further reduce hybridization of the primer to target DNA of a group other than the intended target group. Under certain circumstances, one or more mismatches may be introduced into the 5' end to destabilize internal hairpin loops; such changes are not generally expected to enhance the efficiency of the primer.
The following nucleic acid sequences may be comprised in group-specific untranslated region primers for HLA-A which are specific for the groups as indicated in Table 1.
Table I.
Designation She uence N Tgi Soecifcit_vPo i 'on Il-210mS S' ACC Cgg gAA gCC ggg CCT A 10 et 73-92 3' I S 64 C al.

II-230mS S' ggC Agg TCT CAg CgA Ctg A*Oi,03,11,30102-119 3' 18 60 C

Il-226 S S' CTC TgT ggg gAg AAg CAA A*02 29-47 C 3' 19 60 C

Il-221m11S S' ggg AgC ggC gCC ggg AC 3' A*0301 77-93 II-209 S S' gAA gCA Agg ggC Ccg CCC A10 et 41-S8 18 64 C al.

Il-214mS S' CgC CTg gCg ggg gGg CAA A*230I,24 S4-71 3' 18 66 C

Il-223dS S' gTg AgT gCg ggg TCg Tgg A19 1-19 3' 18 62 C

II-22Sms S' gCC ggg Agg Agg gAC ggT A*30 86-103 3' 18 64 C

II-237m14S S' ggC gCg CCC ggC ggg gA 3' A*29 49-6S

Il-240 S S' ggA ggA ggg Tcg ggC ggA A*31,33 90-107 3' 18 64 C

S'FL-243S S' AgT gTC TTC gCg gTC gCT A*11 S3-71 C 3' 19 62 C

5'FR-2S7S S' CTC AgA TTC TCC CCA gAC Aall, except g 3' 19 60 C for A*

5'FR-273S 5' CAT gCC gAg ggT TTC TCC A*28,6602,6603 CA 3' 20 64 C 360-380 BP202 S S' CTg gCC CTg ACC CAg ACC A*7401,7403Exon 1, A 3' 19 64 C 49-68 BP203 S S' CCT gAC CCA gAC CTg ggC A*8001 Exon l, A 3' 19 64 C SS-73 BP142 AS S' C AGG TAT CTG CGG AGC CCG A*0101/**24227-24S
3' 19 64 C

I3-236 AS S' gTC TgT CAg gAA gAg TCA A*non 02.28584..+2 gAA 3' 21 62 C

S' gTg gAA
AAT
TCT
AgT
CCC
TgA
A3' C
A*multi,notA1,3,11,30.9 I3-246 AS S' AgA TCT ACA ggC gAT CAg A*30 24-43 gA 3' 20 60 C

I3-247m6AS S' gCC AgC CCg ggA gTT CTA A*01,11 36-54 T 3' 19 62 C

I3-249 AS S' CAg AgT CAC TCT CTg gTA
Cag 21 62 C All A,weakS9 J
E
G

I3-280m18AS S' gCg ATC gTC TTC CCg TCA , , C 3' 19 62 C , , , , A*01,03,11,30221-239 I3-282 AS S' AgA gTC ACT CTC Tgg TAC A*8001 148-168 AgA 3' 21 62 C

SUBSTITUTE SHEET ( rule 26 ) The sequences in Table 1 have the following sequence identifiers: Il-210 is SEQ ID N0:35 and the remaining sequences Il-230m through 13-282 have SEQ
ID N0:181-202, respectively.
The present invention provides for nucleic acid molecules comprising regions having the foregoing sequences or their functional equivalents.
"Functional equivalents" of a nucleotide sequence, as defined herein, refers to nucleotide sequences which, when contained in a nucleic acid molecule, retain the specificity of the disclosed sequence and/or hybridize to the complement of the disclosed sequence under stringent hybridization conditions (e.g., .1 x SSC at 65°C).
In specific nonlimiting examples, oligonucleotides comprising the above sequences, or functional equivalents thereof which retain specificity, may be used in a PCR amplif canon reaction in the following pairwise combinations to generate group specific fragments of the lengtl;~s as indicated in Table 2.
SUBSTITUTE SHEET ( rule 26 ) Table 2.
No. Primer Sense PrimerAntisense Size of HLA-A
Mix Primer Product Specificity 1 1.1 I1-230m BP142 785 by A*O1 2 1.2 5'FR-257 I3-247m6 1068 by A*O1 3 1.3 I1-230m I3-247mg 870 by A*Ol,l 1 4 2 I1-226 I3-249 1056 by A*02 3 I1-221m11 I3-280m18 1078 by A*03 6 11 5'FL-243 I3-249 1229 by A* 11 7 9 I1-214m I3-249 1033 by A*23,24 8 10.1 I1-210m I3-236 1450 by A* 10 9 10.2 I1-210m I3-249 1014 by A* 10,68,69 28 5'FR-273 I3-249 1537 by A*68,69, 6602,6603 11 19.1 I1-223d I3-239 1084 by A*29,31,32, or I3- 33,74 12 19.2 Il-240 I3-249 996 by A*31,33 13 29 II-237m14 I3-249 1037 by A*29 14 30 I1-225m I3-249 1000 by A*30 74 BP202 I3-249 1109 by A*7401, (Exon 1 7403 ) 16 80 BP203 I3-282 i 103 by A*8001 (untested) SUBSTITUTE SHEET ( rule 26 ) The following nucleic acid sequences rnay be comprised in group-specific exon region primers for HLA-A which are specific for the groups as indicated in Table 3 (sense primers) and Table 4 (antisens~e primers}. The present invention provides for nucleic acid molecules comprising :regions having the foregoing sequences or their functional equivalents. They may, in specific nonlimiting examples, be used in pairs as set forth in Table :i. The sequences in Table 3, primer numbers 85, 118, 120, 123, 127, 129, 134, 137, 140,160, 167, 175, 193 and 202 have SEQ ID N0:203-216, respectively. The sequences in Table 4, primer numbers 98, 115, 116, 117, 126, 133, 135, 136, 138, 142, 144, 145, 152, 153, 154, 155, 161, 165, 168 and 180, have SEQ ID NU:217-236, respectively, and primer number 119 has SEQ ID N0:245.
SUBSTITUTE SHEET ( rule 26 ) Table3 Primer Localization Sequence Number 85 Exon -4 - 5 5'CTC CTC gTC CCC Agg CTC T 3' 118 Exon 6 - 19 5'TCC ATg Agg TAT TTC TAC ACC 3' 120 Exon -6 - 12 S'ggC CAg gTT CTC AgA CCA 3' 123 Exon 36 - 53 S'CCC ggC CCg gCA gTg gA 3' 127 Exon 1 - 20 5'gTT CTC ACA CCA TCC AgA Tg 3' 129 Exon 4 - 25 5'TCA CAC CCT CCA gAT gAT gTT 3' 134 Exon 63 - 80 5'ggg TAC CAg CAg gAC gCT 3' 137 Exon 9 - 29 5'TCC ATg Agg TAT TTC ACC ACA 3' 140 Exon -1 - 20 5'ggT TCT CAC ACC ATC CAg ATA 3' 160 Exon 1 - 20 5'gTT CTC ACA CCA TCC AgA gg 3' 167 Exon 54 - 71 5'gAg CCC CgC TTC AAC gCC 3' 175 Exon 63 - 71 5'CTT CCT CCg Cgg gTA TgA A 3' 193 Exon 167 - S'gCC ggA gTA TTg ggA CCg 3' 202 Exon 49 - 67 S'CTg gCC CTg ACC CTg ACC A 3' SUBSTITUTE SHEET ( rule 26 ) Table 4.
Antisense primers Primer Localization Sequence Number 98 Exon 2 226 5' gCA ggg TCC CCA ggT CCA 3' 115 Exon 3 195 5' CCT CCA ggT Agg CTC TCA A 3' 116 Exon 3 195 S' CCT CCA ggT Agg CTC TCC A 3' 117 Exon 3 195-213S' CCT CCA ggT Agg CTC TCT g 3' 119 Exon 2 184 5' CTT CAC ATT CCg TgT CTC CT 3' 126 Exon 3 212 5' CCA CTC CAC gCA CgT gCC A 3' 133 Exon 2 229 S' ggA gCg C;gA TCC gcA ggC 3' 135 Exon 3 216 5' ggA gCC ACT CCA Cgg ACC g 3' 136 Exon 3 216 5' gAg CCA CTC CAC gCA CTC 3' 138 Exon 2 186 S' ggC CTT CAC ATT CCg TgT gTT 3' 142 Exon 3 228 5' CAg gTA 'TCT gCg gAg CCC g 3' 144 Exon 2 165 5' Tgg TCC (~AA TAC TCA ggC CT 3' 145 Exon 2 226 5' gCA ggg T'CC CCA ggT TCg 3' 152 Exon 3 163 5' ggg CCg C;CT CCC AgT TgT 3' 153 Exon 2 179 5' TCT gTg AgT ggg CCT aCA CA 3' 154 Exon 2 184 5' CCT TCA CAT TCC gTg TCT gCA 3' 155 Exon 3 216 5' gAg CCA CTC CAC gCA CgT 3' 161 Exon 2 209 5' CCA CTC ggT CAg TCT CTg AC 3' 165 Exon 3 105 5' gAg CgC~, ggT CCT CgT TCA A 3' 168 Exon 2 198 S' gTC TgT ~;Ag Tgg gCC aTC AT 3' 180 Exon 2 12 5' CAg CCA TAC ATC CTC Agg AC 3' SUBSTITUTE SHEET ( rule 26 ) Table 5.
Primer Sense AntisenseSize HLA-A Specificity Mix Primer Primer of No. Product Name 1 1 140 142 247 by A*O10I,0102,800I

2 2 85 98 256 by A*0201-0220 3 3 I40 126 230 by A*0301,0302,0303 4 36 167 168 164 by A*0101,3601 11 I 18 119 195 by A* 1101-1103 6 23 129 115 209 by A*2301 7 24 129 116+117 209 BP A*2402-241 I

8 10.1 160 135 233 by A*2501,2601-2603,2605,4301,6601 9 25 118 133 238 by A*2501,2502 26 l I8 145 235 by A*2601,2602,2604,4301 11 34 134 155 171 by A*3401,3402 12 6602 134 136 240 by A*6602,6603 13 10.2 118 161 222 by A*11,34,6601,6602,68011, 6802,6901 14 43 118 154 196 by A*4301 IS 68 120 152 185 by A*68011,68012,6802,6803 16 69 193 180 375 by A*6901 17 19 127 165 124 by A*2901,2902,310I2,3201, 3301-3303, A*7401-7403 18 29 137 145 236 by A*2901,2902 19 30 175 115+116 162 by A*3001-3002 31 167 144 176 by A*31012 21 32 167 133 159 by A*320I,3202,2501,2502 22 33 137 138 198 by A*3301-3303 23 74 202 153 370 by A*7401,7403 24 80 140 136 234 by A*8001 SUBSTITUTE SHEET ( rule 26 ) In general, the foregoing group-specific primers may be modified by addition, deletion, or substitution of bases, to produce functionally equivalent primers with the substantially the same specificity, that is to say, such that the group specific polymorphism(s) are not removed. Such modij;ications may be constrained by several parameters. First, exact matching at the 3' end is particularly important for primer extension. Preferably, at least 5 nt are complementary to target DNA. When the exactly conserved region is short, for example, less than 10 nt, it is not advisable to change the primer sequences. The primer is preferably less than 50% G or C.
Also, the primers should be designed to avoid specific hybridization with pseudogenes or non-classical HLA Class I loci. In the examples which follow, the melting temperature of all primers used was about 62C to ensure uniform amplification conditions.
For sequencing purposes, the following nucleic acid sequences are sequences which hybridize to all alleles of the indicated loci, in the locations indicated (and hence are referred to as universal sequencing primers).
SUBSTITUTE SHEET { rule 26 ) Table 6:
Universal Sequencing Primers for HLA-A
Desig. Sequence Location Melting Temp.

5'-Ex2(Aw3)5' GCG CCG GGA GGA GGG Int-1 Tm=58-62C
TC 3' 3'-Ex2 S' ATC TCG GAC CCG GAG Int-2 Tm=58C
ACT 3', 5'-Ex3 5' GTT TCA TTT TCA GTT 3' Int-2 Tm=60 C
TAG GCC A

3'-Ex3(Aw6)5' CGG GAG ATC TAC AGG GG 3' Int-3Tm=58-62C
CGA TCA

Desig. Sequence Location Melting Temp.

5'-Ex2(Aw3)5'GCG CCG GGA GGA GGG Int-1 Tm=58-62C
TC 3' 3'-Ex2 5'GTC GTG ACC TGC GCC Int-2 Tm=58-62C
CC 3', 5'-Ex3 5'GGG CGG GGC GGG GCT Int-2 Tm=58-62C
CGG G 3, 3'-Ex3(Aw6)5'CGG GAG ATC TAC AGG Tm=58-62C
CGA TCA GG 3' Int-3 Desig. Sequence Location Melting Temp.

5'-Ex2(Aw3)5' GCG CCG GGA GGA GGG Int-1 Tm=58-62C
TC 3' 3'-Ex2(ABCwl) Int-2 Tm=58-62C
5' GGT
CGT GAC
CT(T/C)CGC
CCC 3' 5'-Ex3(ABCw2) Int-2 Tm=58-62C
5' CCC
GGT TTC
ATT TTC
3' 3'-Ex3(Aw6)5'CGG GAG ATC TAC AGG Tm=58-62C
CGA TCA GG 3' Int-3 The primers in Table 6 are assigned, consecutively, SEQ ID NO: 237-244.
The foregoing three groups of primers include 5' and 3' primers for sequencing across exons 2 and 3, respectively.
The selection of suitable universal sequencing primers is constrained by a variety of rules including the following. Sequencing primer hybridization sites must lie within the fragment amplified by the group specific amplification primers.
All primers are desirably selected to provide informative sequence and not start too far downstream of useful sequence. Preferred primers hybridize to conserved sites near the exon/intron boundaries.
Direct sequencing of the 2nd and 3rd exon may be performed from either the 5' or 3' end using the primers of Table 6 supra which are located in conserved regions of the 1 st, 2nd and 3rd intron as indicated. These conserved regions were found to be identical in all samples investigated, regardless of the amplified group.
An important issue of direct sequencing for HLA class I genes is the generation of a specific PCR product, which is rather complicated due to the SUBSTITUTE SHEET ( rule 26 ) extensive sequence homologies between the different HLA class I loci including several pseudogenes. If an adequate PCR product has been generated, any sequencing chemistry should be applicable.
In the normal case, since group specific amplifications take place before sequencing, only one allele at a time is sequenced, resulting in unambiguous homozygous sequencing results. In these cases alleles are relatively easy to identify, even without software.
However, in about 5% of cases, both alleles come from the same group, but the sequence results show heterozy,gosity. In practice, when viewed by a fluorescence-detecting system, the sample appears as a normal sequence of bases with, sporadically, two bases at one site, each with half the peak height.
This result flows from the high degree of similarity shared among all alleles of each HLA
gene;
sequence heterozygosity flows from base substitutions. The laborious task of determining which alleles are present in the test sequence may be simplified using computer analysis. A software program called GeneLibrarian developed by Visible Genetics, the assignee of the present application, rapidly compares the test sequence to a database which includes all possible homozygote and heterozygote combinations of the alleles. The program identifies those stored sequences that are closest matched to the test sequence. The operator can then determine which allelic pair is in the test sample. If no allelic pair shows an exact match, the software allows the operator to review the test sequence to determine if errors in base-calling or other artifacts are interfering with the analysis.
The order of sequencing reactions may be selected by the operator.
Each exon of each locus may be sequenced on the sense strand or anti-sense strand. A
preferred method is to obtain sequence from one strand from each exon. If the results contain ambiguities, then the amplicon is re-sequenced using the other primer for the same exon. The availability of both sequencing primers provides redundancy to ensure robust results.
In some cases, it may be advantageous to employ an equimolar mixture of 2 or more oligonucleotide species. Mixtures of oligonucleotides may be selected such that between them they will effectively prime the sequencing reactions for all SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCTlCA97/00955 alleles of the locus at the same site.
In an alternative technique, instead of using dye terminators, a dye-labelled primer may be employed. In this case, the selected sequencing primers is labelled on the 5' end with a detectable label, using phosphoramidite or NHS/dye ester techniques well known in the art. The label selected depends on the detection instrument employed. The label for use with an OpenGene System (Visible Genetics Inc., Toronto, ON) is the fluorophore Cy5.5 (Amersham Life Sciences, Cleveland OH). Fluorescein-isothio-cyanate may be used for detection with the ALF
Automated Sequencer (Pharmacia, Piscataway NJ). In this method, which is well known to one skilled in the art, the sequencing reaction mixture is changed slightly to include only one ddNTP per reaction mixture. For detection of reaction products, the sample may be mixed with an equal volume of loading buffer (5% ficoll plus a coloured dye). 1.5 ul of these samples may be loaded per lane of a MicroCel electrophoresis cassette loaded in a MicroGene Blaster automated DNA sequencer (Visible Genetics Inc., Toronto). The sample may be electrophoresed and read.
Results may be displayed and analyzed with GeneObjects software.
The sequence of bases may be determined, and the HLA allele to which the sequence corresponds may then be identified. This process may be performed for each locus (HLA-A, HLA-B, HLA-C) and the results may then be reported to the patient file.
It is well known in the art that different variations of sequencing chemistry may be employed with different automated DNA sequencing instruments.
Single dye instruments, such as the OpenGene System (Visible Genetics Inc., Toronto), the ALF Express (Pharmacia, Uppsala, Sweden) or the Li-Cor 4000L
(Lincoln City, Nebraska) generally use dye-labeled primers. In these systems a single chain termination sequencing reaction mixture is run per lane.
Mufti-dye sequencers, such as the Prism 377 (applied Biosystems, Inc., Foster City, California) detect multiple dyes in a single lane. This technology conveniently employs dye-terminator chemistry, where the chain-terminating nucleotides are themselves labeled with fluorophores (see United States Patent No.
5,332,666, to Dupont de Nemours and Co.). In this case, the reaction products carrying four different labels may be run in a single lane.
SUBSTITUTE SHEET ( rule 26 ) Either single dye or mufti-dye chemistry may be employed according to the present invention, along with other sequencing chemistries. Additional methods for reducing the numbers of reactions required to obtain detailed sequence information from the classical HLA Class I loci are disclosed in commonly owned United States Patent Applications USSN 08/577,858 (for single-track sequencing) and USSN 08/640,672 and 08/684,498 (for single-l:ube sequencing), incorporated by reference herein.
The nucleic acids described above may be comprised in a kit for use in practicing the methods of the invention. In addition to the group-specific primers and primer pairs disclosed, such kits may further comprise buffers, reagents, and enzymes such as, amplification enzymes including but not limited to, Taq polymerase.
In specific, non-limiting embodiments, the kit may comprise group-specific exon region primers (for example, as a "cocktail" comprising a plurality of primers) as well as group-specific untranslated region primers; such primers may be contained in individual tubes.
6.Examg~e~ Determination Of HLA~~roup T~~pe Genomic DNA was prepared firom patient samples according to standard methods, such as a standard salting-out procedure (as provided by the Puregene DNA Isolation Kit, Gentra Systems, Inc., Minneapolis) or by detergent and proteinase K treatment (Current Protocols in Molecular Biology, Eds. Ausubel, F.M.
et al, (John Wiley & Sons; 1995)).
All primers were synthesized on a Gene Assembler plus (Pharmacia, Uppsala, Sweden), and purified by fast protein liquid chromatography. The sequence, length, melting temperature (Tm), group specificity localization of the primers are given in Tables 3 (sense primers), 4 (antisense primers) and S (primer pairs).
Internal positive control primers were: 5' primer hGHI 5'GCC TTC CCA ACC ATT CCC
TTA 3', 2lmer, Tm=64°C, nucleotide position 5560-5580; 3' primer hGHI
5' TCC
ATG TCC TTC CTG AAG CA 3', 20mer, Tm=60°C, nucleotide position 6614-6633.
These control primers amplify a 1074 by fragnnent of the human growth hormone gene.
Group-specific identification was performed as follows. Aliquots of SUBSTITUTE SHEET ( rule 26 ) genomic DNA were separately reacted with a panel of 24 group-specific exon region primer pairs set forth in Table S, supra (see Blasczyk et al., 1995, Tissue Ant. 4f :86-95). An amplification cocktail for pairs of primers was prepared in 10 lcl volume using standard 1 Ox Perkin-Elmer buffer ( 1 x buffer: 50 mM KCI; 1.5 mM MgCl2;
mM Tris-Hcl, pH 8.3; 0.001 % (w/v) gelatin) supplemented with 5% glycerol and 0.1 pl Cresol-red, sodium salt (Cresol-red stock solution:l0 mg/ml). The use of glycerol and cresol red avoids the necessity of using an agarose gel loading buffer.
Additionally, glycerol increases the PCR yield.
The PCR mix for a single SSP tube was as follows:
Genomic DNA 100 ng - 1.00 g I

Taq polymerase, 0.4 - 0.08 pl U

dNTPs, 10 mM - 0.80 ~.1 Buffer, lOx - 1.00 pl Glycerol - 0.50 ~.l Cresol red l Omg/ml - 0.10 ~ l dHzO - 1.52 pl Primer Pair + Control Primer Pair - 5.04 ul Total 10.00 pl The PCR solution was prepared in volumes that would accommodate 30 reactions. The amount of primers used in each 10 pl PCR volume was 3 pmol of each HLA-A primer and 0.8 pmol of each internal control primer.
The reaction mixture was mixed well, then heated in a Thermo-Cycler 9600 (Perkin-Elmer, Inc) and subjected to the following protocol. After an initial denaturation, a first round with 10 two-temperature cycles was followed by 20 three-temperature cycles.
1 ) Initial denaturation at 95°C for 5 min.
2) First 10 cycles i) Denaturation at 95°C for 30 sec.
ii) Annealing and extension at 65°C for 50 sec.
3) Last 20 cycles i) Denaturation at 95°C for 30 sec.
ii) Annealing at 62°C for 50 sec.
iii) Extension at 72°C for 30 sec.
SUBSTITUTE SHEET ( rule 26 ) The reaction tube was then cooled on ice. For visualization, 8 ul of the amplification product were run on a 2 % agaros;e gel prestained with ethidium bromide (0.2 ug/ml). The results were comparc;d to a control lane with known size markers. The reaction products were visualized either as two bands (alleles from different groups) or a single band (alleles from same group). The size of the bands) - were determined and group specificity was assigned according to the length assignments in Table 5.
Figures 10 and 11 show typical gel results, which, as shown in Tables and 11, were interpreted to determine what group specificities were present in genomic DNA samples tested. In Tables 7 and 8, the column titled "Position"
refers to the primer mix no. of Table S.
SUBSTITUTE SHEET ( rule 26 ) Table 7.
PositionHLA S~ecificit3r Kontr. Species .
ampl PM

1 A*0101,0102,8001 1 2 A*0201-0217 2 3 A*0301,0302 3 4 A*0101,3601 36 5 A*1101,1102 11 6 A*2301 23 7 A*2402-2407 24 8 A*2603,2605,6601 X 10.1 9 A*2501 25 10 A*2601,2602,2604,4301 26 11 A*3401,3402 34 12 A*6602 6602 13 A*1101,1102,3401,3402, X 10.2 6601,6602, A*68011,6802,6901 14 A*4301 43 15 A*68012,6802,6803 68 16 A*6901 69 17 A*2901,2902,3101,3201 X 19 3301-3303, A*7401 18 A*2901,2902 29 19 A*3001-3004 30 20 A*3101 31 21 A*3201,2501 X 32 SUBSTITUTE SHEET ( rule 26 ) Table 8.
Position HLA Specificity Kontr. S~ecie~s Am~l. PM
1 A*0101,0102,8001 I

- 2 A*0201-0217 %'~ 2 3 A*0301,0302 3 4 A*0101,3601 36 A*1101,1102 11 6 A*2301 23 7 A*2402-2407 24 8 A*2501,2601-2603, 10.1 2605,6601 A*2501 25 A*2601,2602,2604,4301 26 11 A*3401,3402 34 12 A*6602 6602 13 A*1101,1102,3401,3402 :~ 10.2 6601,6602, A*68011,6802, 14 A*4301 43 A*6801,6802 :K 68 16 A*6901 69 17 A*2901-2902,3101,3201, 19 3301-3303, A*7401 18 A*2901,2902 29 19 A*3001-3004 30 SUBSTITUTE SHIEET ( rule 26 ) 7. Example: Determination Of Group Specificity Using A Primer Cocktail Group specific low-resolution typing of the patient sample may be performed as follows. First, a stock PCR amplification reaction mixture may be prepared for 30 reactions:
~_1 dNTPs IOmM 24 Glycerol 100% 15 l Ox PCR Buffer* 30 Cresol-red ( 1 Omg/ml) 3.0 final 117 *1 X PCR Buffer comprises 10 MM Tris-HCl pH 8.3, 50 mM KCI, 1.5 mM MgCl2 and 0.001 % (w/v) gelatin.
The stock mixture may be prepared in a large volume and be stored for at least one month at 4 ° C or be aliquoted ( 117.0 ~l) and stored at -3 0 ° C for at least six months. Repeated thawing and freezing should be avoided.
A mixture containing all the HLA-A group specific amplification primers listed in Table 5 may be prepared separately (the "Cocktail"). One member of each primer pair is labelled on the 5' end with a fluorescent label. Final Cocktail concentrations may be designed to provide 3 pmol of each HLA-A primer per 5 ul.
Optionally, an internal control primer may be added (to determine among other things, the success of amplification) in the amount of 0.8 pmol per S ul.
Suitable internal control primers amplify a 1074 by fragment of the human growth hormone gene (see supra).
SUBSTITUTE SHEET ( rule 26 ) To perform the low resolution amplification reaction, the reaction mixture may be prepared as follows:
Volume Stock Mixture 5 ~1 Cocktail S ~1 Patient sample DNA 100-250 ng 1 ~1 Taq Polymerase Enzyme 0.4 U 0.08 ~1 PCR cycle parameters may be adjusted for a Perkin-Elmer System 9600 thermal cycler. After an initial denaturation, a first round with 10 two-temperature cycles may be followed by 20 three-temperature cycles: 1 ) Initial Denaturation at 95 ° C for 5 min; 2) First 10 cycles i) Denaturation at 95 ° C for 30 seconds and ii) Annealing and extension at 65 °C for 50 seconds; 3) Last 20 cycles i) Denaturation at 95°C for 30 seconds, ii) Annealing at 62°C for SO seconds and iii) Extension at 72°C for 30 seconds.
The reaction tube may then be cooled on ice. For visualization, 2ul of the amplification product may be run on a polyacrylamide gel giving single nucleotide length resolution such as in a MicroGene Blascter. The results were compared to a control lane with known size markers. The reaction products may be visualized either as two bands (alleles from different groups) or a single band (alleles from same group). The size of the bands) may be determined and group specificity may be assigned according to the length assignments in Table S.
SUBSTITUTE SHEET ( rule 26 ) 8. Example- Determination Of Allelic Type By Sequencing After determining group type specificity, group specific amplification of a fresh portion of the patient sample may be performed using a single pair of primers specific for the group in question to generate sequencing template. In a pre-ferred, nonlimiting embodiment, amplification primers may be selected from Table 2, supra, which lists group-specific untranslated region primers. This second amplification serves two purposes. First, it confirms, by successful amplification, the group determination of the low resolution test. Second, it generates sequence information which may be used for accurate allele identification. If two groups are identified, two separate reactions may be performed each using a different primer pair.
8.1. PCR Protocol The same PCR protocol may be used for all primer mixes used for template generation. The PCR amplification may be set up in a total volume of SOuI
in order to produce enough PCR product for more than 10 sequencing reactions.
This ensures that, if anything fails during the sequencing process, sequencing can be repeated without generation of a new template. The high stringency of the PCR
primers and protocol detailed below makes the use of a "hot start approach"
unnecessary. The following PCR reaction mix may be used:
SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCT/CA9?/00955 volume nor reaction SX PCR buffer* 10.0,u1 DMSO l.O~c1 2.SmM each dNTP S.O~cI

ddH20 27.

_ 43.8~c1 Total - Sense primer** (lOpmol/~cI) 1.0~c1 Antisense primer**(lOpmol/~cI) l.O~c1 Taq Polymerase (SU/~l) 0.2~c1 Genomic DNA ( 1 OOng/~cl) 4.Ou1 Final Total 50.01 *Composition of 5X PCR buffer:75mDd (NH4)2504; l7.SmM MgCl2; and 300mM Tris-HCL, pH 9.0 **The pair of group specific amplification primers may be selected from those disclosed in Table 2, supra.
PCR cycle parameters may be .adjusted for a Perkin-Elmer System 9600 thermal cycler. After an initial denaturation, a first round with 10 two-temperature cycles may be followed by 20 three-temperature cycles.
1.) Initial Denaturation at 95 C for 5 min 2.) First 10 cycles i) Denaturation at 95 C for 30 seconds ii) Annealing and extension at 65 C for 50 seconds 3.) Last 20 cycles i) Denaturation at 95 C for 30 seconds ii) Annealing at 62 C for :i0 seconds iii) Extension at 72 C for ?.0 seconds l0,ul of the PCR product may then be run on a 2 % agarose gel prestained with ethidium bromide (0,2 ~g/ml). A distinct band of the expected size should be seen.
8.2. Sequencing Reaction Protocol The sequencing reactions may be carried out with AmpliTaqTM DNA
Polymerase FS dye terminator cycle sequencing chemistry using the Ready Reaction DyeDeoxy Terminator Cycle Sequencing Kit FS (Perkin Elmer Applied Biosystems SUBSTITUTE SHEET ( rule 26 ) Division, Foster City, CA) according to the manufacturer's protocol. This Kit contains the four ddNTPs with different fluorescence labels (=Dye Terminators). The PCR
fragments may be used directly for sequencing without any prior purification step.
To simplify the pipetting steps, a master mix may be prepared consisting of the 5'Biotin labeled sequencing primer, ddH20 and the Kit reagents.
This master mix should he prepared immediately prior to use and can be kept at room temperature until use. The sequencing master mix for one reaction may comprise 3.0 ~l of a lpmol/~l solution of sequencing primer; 6.0 ~l ddHzO, and 8.0 ~.1 of premixed sequencing reagents; for 36 + 1 reactions, these amounts are increased, respectively, to 111.0 ~1; 222.0 ~1; and 296.0 ~1, respectively. The sequencing primer may be selected from the sequencing primers for HLA-A set forth in Table 6, supra.:
The master mix may be aliquoted in a volume of 171 for each sequencing reaction in a 200,u1 PCR tube and 3~1 of the unpurified PCR product are added. The reaction mixes may then be subjected to 25 cycles in a Perkin Elmer thermal cycler 9600. Each cycle consists of 10 sec 95 C, 5 sec 50 C and 4 min 60 C.
8.3. Purification Of Extension Products After the sequencing reaction the extension products are desirably separated from the unincorporated Dye Terminators which would otherwise interfere with the fluorescence-based detection process of the electrophoretically separated sequencing fragments.
For each sequencing reaction, 50 ~g (5 ~1) Streptavidin-coated Dynabeads M-280 (Dynal Inc., Oslo, Norway) may be washed in 5 ~l of 2x Binding and Washing buffer {"B&W"; 2X B&W buffer: 2M NaCI, IOmM Tris-HC1 pH 7.5, 1mM EDTA). The beads may then be resuspended in 20 p.1 of 2x B&W.
To each 20u1 sequencing reaction, 20,u1 of resuspended beads may be added, and the mixture may be incubated at room temperature (20-25 C) for 15 minutes. The beads may then be immobilized, the supernatant may be removed, and then the beads may be washed once in 70% ethanol by pipetting up and down five times. Then, as much as possible of the ethanol may desirably be removed, because residual ethanol may interfere with electrophoretic gel mobility.
For each sequencing reaction, 4~1 of loading buffer (5:1 Formamide-SUBSTITUTE SHEET ( rule 26 ) 25mM EDTA pH 8.0, SOmg/ml Dextran Blue) may be added.
8.4. Electrophoresis ~r~d Data Collection Samples prepared by the foregoing methods may be used immediately or be stored at 4 C at least for 24 hours before starting the electrophoretic separation.
Prior to the electrophoretic separation, each reaction may be incubated at 90 C for 2 minutes. 3~c1 of each sample may be loaded on a prerun sequencing gel. For an automated ABI 377 sequencer (Applied Biosystems, Foster City, CA) a 0,2mm thick 5% polyacrylamide (acrylamide:bisacrylamide: = 29:1 ) - 7 M urea gel may be used [gel composition: 21.0 g urea, 8.4 ml 30% acr~lamide (stock solution: 58g acryl-amide, 2g bisacrylamide in bidistilled water), 6.0 ml TBE buffer ( 1 Ox TBE-buffer:
108.0 g tris base, 55.0 g boric acid, 7.4 g Na2I:DTA), 15 gel TEMED, 350 ~cl 10%
Ammoniumpersulfate {1.0 g Ammoniumpersulfate in 10 ml ddH20), 20.0 mI
ddHZO]. Electrophoresis may be run at constant 48 watt for 8h. Data collection may be initiated immediately after starting the electrophoresis on the ABI377. Data analyses may be perfonmed thereafter using the ABI analysis software (version 2.1.1 ).
8.5. Data Internretation And HLA Ty~~~~ng After data collection, the chromatograms may be printed and sequences may be compared manually to existing HLA data in the EMBL databank and the sequences compiled by Arnett and Pa:rham. Due to the group-specific amplification and the lack of heterozygous positions, manual analysis is typically very fast. Alternatively, sequences may be checked with the data analysis editor (Sequence NavigatorTM, Applied Biosystems) and aligr.~ed with any sequence alignment program.
Various publications are cited herein, the contents of which are hereby incorporated by reference in their entireties.
SUBSTITUTE SHEET ( rule 26 ) SEQUENCE LISTING
(1) GENERAL INFORMATION
(i) APPLICANT:
(ii) TITLE OF THE INVENTION: Method and Kit for HLA Class I Typing (iii) NUMBER OF SEQUENCES: 245 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Baker & Botts, L.L.P.
(B) STREET: 30 Rockefeller Plaza (C) CITY: New York (D) STATE: NY
(E} COUNTRY: U.S.A.
(F) ZIP: 10112-0228 (v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Diskette (B) COMPUTER: IBM Compatible (C) OPERATING SYSTEM: ASCII DOS text (D) SOFTWARE: FastSEQ Version 2.0 (vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 08/766,189 (B) FILING DATE: 12 DEC 1996 (viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Tenser, Arthur and Kole, Lisa (B) REGISTRATION NUMBER: 18,839 and 35,225 (C) REFERENCE/DOCKET NUMBER: 30861 (ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: 212-705-5000 (B) TELEFAX: 212-705-5020 (C) TELEX:
(2) INFORMATION FOR SEQ ID NO:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single SUBSTITUTE SHEET ( rule 26 ) (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A 5' Flanking Region Allele consensus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCACCiCACGCCTGAAATCTTGGCGCTGG 60 CGCTGCTGTGACTAACCGAAGAGACCTTTGGGCTGTCiGGTTACCCTCACTCTTGACCCAG 120 GCGCAGCACTCATAGGTCCTTCTTCCTGGGATGTATC:CAACCCTCTCCCTCTTTTCTTTG 180 ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGG'.CTAGGTATGTGGCGGAAGCCCCGG 240 CCCTCTTTTGGGAGCCGTACCCGGGGCAGGGAGAGGi~AP.GTGAAAAGTAGGGCATTAGAG 420 (2) INFORMATION FOR SEQ ID N0::2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 0101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCAC'GCACGCCTGAAATCTTGTCGCTGG 60 CGCTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC'.CGGGTCCTCGACAGACTCTTTGTT 360 (2) INFORMATION FOR SEQ ID N0:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear SUBSTITUTE SHEET ( rule 26 ) (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
{A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 0301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:3:

(2) INFORMATION FOR SEQ ID N0:4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human fix) FEATURE:
(A) NAME/FCEY: HLA-A 5' Flanking Region Allele A* 1101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4:

(2) INFORMATION FOR SEQ ID N0:5:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no SUBSTITUTE SHEET ( rule 26 ) (iv) ANTI-SENSE: no (v) ORIGINAL 50URCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A*1102 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:5:

CGCTTCTGTG ACTAACCGAAGAGACCTTTGGGCTGTGiGGTTACCCTCACTCTTGACCCAG 120 GCGCAGCACT CATAGGTCCTACTTCCTGGGATGCATC'.CAACCCTCTCCCTCTTTTCTTTG 180 CGCTTCGCGA GACAGAGTTACAGAGGGACTCAGAACC:GGGTCCTCGACAGACTCTTTGTT 360 (2) INFORMATION FOR SEQ ID N0:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B} TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:6:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCAC'GCACGCCCGAAATCTTGTCGCTGG 60 ACTCTGGGACTCTCGGTGCGGACCCCGGGACCCTGF.AGCGGGACTGGGGAGACGAGGACA 300 CGCTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC'.CGGGTTCTCGACAGACTCTTTGTT 360 (2) INFORMATION FOR SEQ ID N0:7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHF;ET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3002 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:7:

(2) INFORMATION FOR SEQ ID N0:8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL 50URCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3004 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:8:

(2) INFORMATION FOR SEQ ID N0:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 0201-SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:9:

GCGCAGCACTCATAGGTCCTACTTCCTGGGATGTATC'.CAACCCTCTCCCTCTTTTCTTTG 180 ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGG7.'TAGGTATGTGGCGGAAGCCCCGG 240 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAACC:GGGTTCTCGACAGACTCTTTGTT 360 (2) INFORMATION FOR SEQ ID N0::10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 0215 (xi) SEQUENCE DESCRIPTION: SEQ ILK N0:10:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCAC:GCACGCCTGAAATC TTGGCGCTGG60 CGCTGCTGTGACTAACCGAAGAGACCTTTGGGCTG7.'GGGTTACCCTCACT CTTGACCCAG120 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC:CGGGTTCTCGACAG ACTCTTTGTT360 (2) INFORMATION FOR SEQ ID N0:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 0217 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:11:

(2) INFORMATION FOR SEQ ID N0:12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6801 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:12:

(2) INFORMATION FOR SEQ ID N0:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6802 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:13:
SUBSTITUTE SHEET ( rule 26 ) ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGGT'TAGGTATGTGGCGGAAGCCCCGG 240 ACTCTGGGACTCTCGGTGCGGACCCCGGGACCCTGAP,GCGGGACTGGGGAGACGAGGACA 300 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAACC'.GGGTTCTCGACAGACTCTTTGTA 360 CCCTCTTTGGGAGCCGTACCCGGGGCAGGGAGAGGAP,AGTGAAAAATAGGGCATTAGAGA 420 (2) INFORMATION FOR SEQ ID N0:7.4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:

GCGCAGCACTCATAGGTCCTTCTTCCTGGGATGTAT'CCAACCCTCTCCCTCTTTTCTTTG180 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC'CGGGTTCTCGACAGACTCTTTGTA360 CCCTCTTTGGGAGCCGTACCCGGGGCAGGGAGAGGF,AAGTGAAAAATAGGGCATTAGAGA420 (2) INFORMATION FOR SEQ ID N0:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2301 (xi) SEQUENCE DESCRIPTION: SEQ II) N0:15:

CGCTGCTGTG ACTAACCGAA GAGACCTTTG GGCTG'CGGGT TACCCTCACT CTTGACCCAG 120 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ICEY: HLA-A 5' Flanking Region Allele A* 2402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:16:

(2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:17:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear {ii) MOLECULE TYPE: genomic: DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2404 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:18:

ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGG'TTAGGTATGTGGCG GAAGCCCCGG240 CCCTCTTTTGGGAGCCGTACCCGGGGCAGGGAGAGG.AAAGTGAAAAGTAG GGCATTAGAG420 (2) INFORMATION FOR SEQ ID N0:19:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomi.c DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2405 (xi) SEQUENCE DESCRIPTION: SEQ III N0:19:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCAC:GCACGCCTGAAATCTTGTCGCTGG60 CGCTGCTGTGACTAACCGAAGAGACCTTTGGGCTG'TGGGTTACCCTCACTCTTGACCCAG120 GCGCAGCACTCATAGGTCCTTCTTCCTGGGATGTA'TCCAACCCTCTCCCTCTTTTCTTTG180 ACTCTGGGACTCTCGGTGCGGACCCCGGGACCCTG;~1AGCGGGACTGGGGAGACGAAGACA300 CGGTTCGCGAGACAGAGTTACAGAGGGACTTAGAA~~CGGGTTCTCGACAGACTCTTTGTT360 SUBSTITUTE SHF;ET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2407 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:

(2) INFORMATION FOR SEQ ID N0:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2501 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:

SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCT/CA97l00955 (2) INFORMATION FOR SEQ ID N0:22:
(i} SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:

ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGG'.CTAGGTATATGGCG GAAGCCCCGG240 (2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:

(2) INFORMATION FOR SEQ ID N0:24:
SUBSTITUTE SHEET ( rule 2G ) $0 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 4301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:

(2) INFORMATION FOR SEQ ID N0:25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:25:

(2) INFORMATION FOR SEQ ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs SUBSTITUTE SHEET ( rule 26 ) (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6602 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:26:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCACG:CACGCCTGAAATCTTGGCGCTGG 60 GCGCAGCACTCATAGGTCCTTCTTCCTGGGATGTATC'.CAACCCTCTCCCTCTTTTCTTTG 180 ACTCTGGGACTCTCGGTGCGGACCCCGGGACCCTGAF.GCGGGACTGGGGAGACGAGGACA 300 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAACC'.GGGTTCTCGACAGACTCTTTGTA 360 CCCTCTTTGGGAGCCGTACCCGGGGCAGGGAGAGGAAP.GTGAAAAATAGGGCATTAGAGA 420 ( 2 ) INFORMATION FOR SEQ ID NO : :? 7 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 6603 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:27:

CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC'CGGGTTCTCGACAG ACTCTTTGTA360 CCCTCTTTGGGAGCCGTACCCGGGGCAGGGAGAGGF,AAGTGAAAAATAGG GCATTAGAGA420 (2) INFORMATION FOR SEQ ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCTlCA97/00955 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:28:

(2) INFORMATION FOR SEQ ID N0:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 2902 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:29:

(2) INFORMATION FOR SEQ ID N0:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B} TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear {ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 31012 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:30:

ACGCCTCAACCCCTTAGGGGTTCCGACCCTGAGGGG'rTAGGTATGTGGCG GAAGCCCCGG240 (2) INFORMATION FOR SEQ ID N0:31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A ~~~ Flanking Region Allele A* 3201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:31:
GAGCCGCAGACCCCTCTTAGACTCAGGGCCACCCA(:GCACGCCTGAAATCTTGGCGCTGG 60 CGCTGCTGTGACTAACCGAAGAGACCTTTGGGCTG".CGGGTTACCCTCACTCTTGACCCAG 120 GCGCAGCACTCATAGGTCCTTCTTCCTGGGATGTA'.CCCAACCGTCTCCCTCTTTTCTTTG 180 ACTCTGGGACTCTCGGTGCGGACCCCGGGACCCTGP.AGCGGGACTGGGGAGACGAGGACA 300 CGGTTCGCGAGACAGAGTTACAGAGGGACTCAGAAC:CGGGTTCTCGACAGACTCTTTGTT 360 (2) INFORMATION FOR SEQ ID N0:32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genom:ic DNA
(iii) HYPOTHETICAL: no (iv} ANTI-SENSE: no SUBSTITUTE SH~:ET ( rule 26 ) (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:32:

(2) INFORMATION FOR SEQ ID N0:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 450 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 3303 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:33:

(2) INFORMATION FOR SEQ ID N0:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHEET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 7401 (xi) SEQUENCE DESCRIPTION: SEQ ID 1J0:34:
w GAGCCGCAGA CCCCTCTTAGACTCAGGGCCACCCACGCACGCCTGAAATC TTGGCGCTGG60 (2) INFORMATION FOR SEQ ID N0:35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE;
(A) NAME/KEY: I1-210m (xi) SEQUENCE DESCRIPTION: SEQ ID N0:35:

(2) INFORMATION FOR SEQ ID N0:36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genom__c DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A S' Flanking Region Allele A* 7402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:36:

SUBSTITUTE SHF;ET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid {C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 7403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:37:

(2) INFORMATION FOR SEQ ID N0:38:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 449 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A 5' Flanking Region Allele A* 8001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:38:

SUBSTITUTE SHEET ( rule 26 ) $7 CCCTCTTTGG GAGCCGTACC CGGGGCAGGG AGAGGAAi~GT GAAAAGTAGG GCATTAGAGA 420 (2) INFORMATION FOR SEQ ID N0:39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FtEY: HLA-A Intron 1 Allele consensus (xi) SEQUENCE DESCRIPTION: SEQ ID N0:39:
GTGAGTGCGG GGTCGGGAGG GAAACGGCCT CTGTGG(3GAG AAGCAAGGGG CCCGCCTGGC 60 (2) INFORMATION FOR SEQ ID N0:40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ItEY: HLA-A 5' Intron Allele A* 0101 (xi) SEQUENCE DESCRIPTION: SEQ ILt N0:40:

GGGGGCGCAG GACCGGGGGA GCCGCGCCGG GAGGAC:GGTC GGGCAGGTCT CAGCCACTGC 120 (2) INFORMATION FOR SEQ ID N0:41:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genom:ic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:41:

(2) INFORMATION FOR SEQ ID N0:42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 1101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:42:

(2) INFORMATION FOR SEQ ID N0:43:
(1) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 1102 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:43:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:4EE:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 230 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear _ (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In.tron 1 Allele A* 3001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:44:

TCGCCCCCAG
(2) INFORMATION FOR SEQ ID NO:~IS:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3002 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:45:
GTGAGTGCGG GGTCGGGAGG GAAACCGCCT CTGCGG,GGAG AAGCAAGGGG CCCTCCTGGC 60 (2) INFORMATION FOR SEQ ID N0:46:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genom~~c DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no SUBSTITUTE SHEET ( rule 26 }

(v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3004 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:46:

(2) INFORMATION FOR SEQ ID N0:47:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:47:

(2) INFORMATION FOR SEQ ID N0:48:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0202 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:48:

CGTCCCCAG

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:49:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A Intron 1 Allele A* 0203 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:49:
GTGAGTGCGG GGTCGGGAGG GAAACGGCCT CTGTGG(3GAG AAGCAACGGG CCGCCTGGCG 60 (2) INFORMATION FOR SEQ ID NO: SO:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A I:ntron 1 Allele A* 0204 (xi) SEQUENCE DESCRIPTION: SEQ II) N0:50:

(2) INFORMATION FOR SEQ ID N0:51:
(i} SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv} ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHF;ET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0205 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:51:

(2) INFORMATION FOR SEQ ID N0:52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0206 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:52:

(2) INFORMATION FOR SEQ ID N0:53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0207 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:53:

(2) INFORMATION FOR SEQ ID N0:54:
SUBSTITUTE SHEET ( rule 26 ) (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0207 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:54:
GTGAGTGCGG GGTCGGGAGG GAAACGGCCT CTGTGGCiGAG AAGCAACGGG CCGCCTGGCG 60 (2) INFORMATION FOR SEQ ID N0:55:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv} ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0208 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:55:

(2) INFORMATION FOR SEQ ID N0:56:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genom3.c DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A :=ntron 1 Allele A* 0209 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:56:

{2) INFORMATION FOR SEQ ID N0:57:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs {B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0210 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:57:

(2) INFORMATION FOR SEQ ID N0:58:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 0211 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:58:

(2) INFORMATION FOR SEQ ID N0:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE. nucleic acid SUBSTITUTE SHEET ( rule 26 ) (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In.tron 1 Allele A* 0215 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:59:

( 2 ) INFORMATION FOR SEQ ID NO : E> 0 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomi~~ DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no {v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
{A) NAME/KEY: HLA-A I:ntron 1 Allele A* 0217 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:60:

(2) INFORMATION FOR SEQ ID N0:61:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STR.ANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no {iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human ' (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6801 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:61:
SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:62:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear {ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6802 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:62:

(2) INFORMATION FOR SEQ ID N0:63:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:63:

(2) INFORMATION FOR SEQ ID N0:64:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs {B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:64:

(2) INFORMATION FOR SEQ ID N0:65:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic: DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:65:

(2) INFORMATION FOR SEQ ID N0:66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:66:

SUBSTITUTE SHF;ET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A} ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2404 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:67:

(2) INFORMATION FOR SEQ ID N0:68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
{A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2405 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:68:

(2) INFORMATION FOR SEQ ID N0:69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no SUBSTITUTE SHEET ( rule 26 ) (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 1 Allele A* 2407 (xi) SEQUENCE DESCRIPTION: SEQ ID PJ0:69:

(2) INFORMATION FOR SEQ ID N0:70:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: i29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2501 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:70:

(2) INFORMATION FOR SEQ ID N0:71:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic: DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix} FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2601 (xi) SEQUENCE DESCRTPTION: SEQ ID N0:71:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:72:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:72:

(2) INFORMATION FOR SEQ ID N0:73:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:73:

(2) INFORMATION FOR SEQ ID N0:74:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHEET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6602 (xi) SEQUENCE DESCRIPTION: SEQ ID Ni0:74:
GTGAGTGCGG GGTCGGGAGG GAAACGGCCT CTGTGGGGiAG AAGCAAGGGG CCCGCCCGGC 60 GGGGGCGCAG GACCCGGGAA GCCGCGCCTG GAGGAGGC~TC GGGCGGGTCT CAGCCACTCC 120 (2) INFORMATION FOR SEQ ID N0:7~i:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: I30 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 6603 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:75:

(2) INFORMATION FOR SEQ ID N0:76:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 230 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic: DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 430I
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:76:

(2) INFORMATION FOR SEQ ID N0:77:
SUBSTITUTE SHEET ( rule 26 ) (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:77:

(2) INFORMATION FOR SEQ ID N0:78:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 2902 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:78:

(2) INFORMATION FOR SEQ ID N0:79:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3/01 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:79:

GGGGCGCAGG ACCCGGGTAG CCGCGCCGGG AGGAGGG'.CCG GGCGGATCTC AGCCACTCCT 120 (2) INFORMATION FOR SEQ ID NO: BO:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In.tron 1 Allele A* 3201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:80:

GGGGCGCAGG ACCCGGGTAG CCGCGCCGGG AGGAGGC~TCG GGCGGGTCTC AGCCACTCCT 120 (2) INFORMATION FOR SEQ ID NO:E31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic~ DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:81:

(2) INFORMATION FOR SEQ ID N0:82:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid SUBSTITUTE SHEET ( rule 26 ) (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii} HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 3303 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:82:

(2) INFORMATION FOR SEQ ID N0:83:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STR.ANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 7401 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:83:

(2) INFORMATION FOR SEQ ID N0:84:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 7402 (xi) SEQUENCE DESCRIPTION. SEQ ID N0:84:
SUBSTITUTE SHEET ( rule 26 ) GTGAGTGCGG GGTCGTGGGG AAACCGCCTC TGCGGGGi~GA AGCAAGGGGC CCGCCCGGCG 60 GGGGCGCAGG ACCCGGGTAG CCGCGCCGGG AGGAGGG'PCG GGCGGGTCTC AGCCACTCCT 120 (2) INFORMATION FOR SEQ ID N0:85:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 129 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ICEY: HLA-A Intron 1 Allele A* 7403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:85:

(2) INFORMATION FOR SEQ ID N0:86:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 130 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 1 Allele A* 8001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:86:

(2) INFORMATION FOR SEQ ID N0:87:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHF;ET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele consensus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:

(2) INFORMATION FOR SEQ ID NO:88:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTO-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0101 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:

(2) INFORMATION FOR SEQ ID NO: 89:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTO-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:89:

CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'rCAGTTTAGGCCAAAAATCCCC180 CCAGGTTGGTCGGGGCGGGGCGGGGCTCGGGGGACCGGt3CTGACCGCGGGGTCCGGGCCA240 G

(2) INFORMATION FOR SEQ ID N0:90:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron Allele A* 0202 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:90:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCTC'rCATCCCCCACGGACGGGCCAGG60 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'TTCAGTTTAGGCCAAAAATCCCC180 G

(2) INFORMATION FOR SEQ ID N0:91:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron Allele A* 0203 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:91:

TCGCCCACAGTCTCCGGGTCCGAGATCCGCCCCGAAt3CCGCGGGACCCCGAGACCCTTGC120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCAT'rTTCAGTTTAGGCCAAAAATCCCC180 CCAGGTTGGTCGGGGCGGGGCGGGGCTCGGGGGACCt3GGCTGACCGCGGGGTCCGGGCCA240 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR 5EQ ID N0:92:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0204 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:92:

(2) INFORMATION FOR SEQ ID N0:93:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0205 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:93:

(2) INFORMATION FOR SEQ ID N0:94:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 2 Allele A* 0206 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:94:

G

(2) INFORMATION FOR SEQ ID N0:95:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii} MOLECULE TYPE: genomic DNA
(iii) HYPOTHETTCAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 2 Allele A* 0207 (xi) SEQUENCE DESCRIPTION: SEQ ID td0:95:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCTC'.CCATCCCCCACGGACGGGCCAGG60 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'.CTCAGTTTAGGCCAAAAATCCCC180 (2) INFORMATION FOR SEQ ID N0:96:
(i} SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0208 SUBSTITUTE SHEET { rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:96:

(2) INFORMATION FOR SEQ ID N0:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0209 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:97:

(2) INFORMATION FOR SEQ ID N0:98:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ICEY: HLA-A Intron 2 Allele A* 0210 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:98:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:9S>:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no _ (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Inl:ron 2 Allele A* 0211 (xi) SEQUENCE DESCRIPTION: SEQ ID 240:99:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCTC'.L'CATCCCCCACGG ACGGGCCAGG60 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'CTCAGTTTAGGCC AAAAATCCCC180 (2) INFORMATION FOR SEQ ID NO:1!)0:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In1_ron Allele A* 0215 (xi) SEQUENCE DESCRIPTION: SEQ ID 1J0:100:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCTC'rCATCCCCCACGGACGGGCCAGG 60 TCGCCCACAGTCTCCGGGTCCGAGATCCGCCCCGAAG(~CGCGGGACCCCGAGACCCTTGC 120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'rTCAGTTTAGGCCAAAAATCCCC 180 CCAGGTTGGTCGGGGCGGGGCGGGGCTCGGGGGACCG(3GCTGACCGCGGGGTCCGGGCCA 240 (2) INFORMATION FOR SEQ ID NO:101:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs . (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no !v) ORIGINAL SOURCE:
(A) ORGANISM: human !ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0217 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:

(2) INFORMATION FOR SEQ ID N0:102:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 6801 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:102:

G

!2) INFORMATION FOR SEQ ID N0:103:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 6802 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:103:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCCCZ'CATCCCCCACGGACGGGCCAGG 60 TCGCCCACAGTCTCCGGGTCCGAGATCCGCCCCGAAGC'CGCGGGACCCCGAGACCCTTGC 120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT7.'TCAGTTTAGGCCAAAAATCCCC 180 (2) INFORMATION FOR SEQ ID N0:104:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int_ron 2 Allele A* 6901 (xi) SEQUENCE DESCRIPTION: SEQ ID iJ0:104:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCTC'fCATCCCCCACGGACGGGCCAGG 60 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'fTCAGTTTAGGCCAAAAATCCCC 180 CCAGGTTGGTCGGGGCGGGGCGGGGCTCGGGGGACCGt3GCTGACCGCGGGGTCCGGGCCA 240 (2) INFORMATION FOR SEQ ID NO:11)5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In~~ron 2 Allele A* 2501 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:105:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCCC'rCATCCCCCACGGACGGGCCAGG 60 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT'TTCAGTTTAGGCCAAAAATCCCC 180 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:106:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:106:

(2) INFORMATION FOR SEQ ID N0:107:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 4301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:107:

(2) INFORMATION FOR SEQ ID NO:108:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 6601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:108:

CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'TCAGTTTAGGCCAAAAATCCCC 180 (2) INFORMATION FOR SEø ID N0:109:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 6602 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:109:

CCCGGGAGAG GCCCAGGCGC CTTTACCCGG TTTCATTT'TC AGTTTAGGCC AAAAATCCCC 180 (2) INFORMATION FOR SEQ ID NO:110:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no - (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 6603 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:

(2) INFORMATION FOR SEQ ID NO:111:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3402 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:

(2) INFORMATION FOR SEQ ID N0:112:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:112:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:113:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic :DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2902 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:113:

(2) INFORMATION FOR SEQ ID N0:114:
(i.) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ICEY: HLA-A Int.ron 2 Allele A* 3101 (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:114:

TCACCCACAGTCTCCGGGTCCGAGATCCACCCCGAAGC'.CGCGGGACCCCGAGACCCTTGC 120 (2) INFORMATION FOR SEQ ID N0:11.5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:115:

(2) INFORMATION FOR SEQ ID N0:116:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:116:

(2) INFORMATION FOR SEQ ID N0:117:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3303 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:117:

CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'.CCAGTTTAGGCCAAAAATCCCC180 (2) INFORMATION FOR SEQ ID N0:118:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic :DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 7401 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:118:

(2) INFORMATION FOR SEQ ID N0:119:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 7402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:119:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:120:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 7403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:120:

(2) INFORMATION FOR SEQ ID N0:121:
(i) SEQUENCE CHARACTERISTICS:
(A} LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:121:

(2) INFORMATION FOR SEQ ID N0:122:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2402 (xi) SEQUENCE DESCRIPTION: SEQ ID Tf0:122:
GTGAGTGACC CCGGCCCGGGGCGCAGGTCACGACCCC7.'CATCCCCCACGGACGGGCCGGG 60 TCGCCCACAG TCTCCGGGTCCGAGATCCACCCCGAAGC:CGCGGGACCCCGAGACCCTTGC 120 CCCGGGAGAG GCCCAGGCGCCTTAACCCGGTTTCATT7.'TCAGTTTAGGCCAAAAATCCCC 180 (2) INFORMATION FOR SEQ ID N0:123:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2403 (xi) SEQUENCE DESCRIPTION: SEQ ID a~0:123:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCCC'TCATCCCCCACGGACGGGCCGGG 60 CCCGGGAGAGGCCCAGGCGCCTTAACCCGGTTTCATT'TTCAGTTTAGGCCAAAAATCCCC 180 (2) INFORMATION FOR SEQ ID N0:124:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2404 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:124:

{2) INFORMATION FOR SEQ ID N0:125:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2405 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:125:

(2) INFORMATION FOR SEQ ID N0:126:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 2407 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:126:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:127:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 0301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:127:

CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'TCAGTTTAGGCCAAAAATCCCC180 (2) INFORMATION FOR SEQ ID N0:128:
(i) SEøUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D} TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 1101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:128:
GTGAGTGACCCCGGCCCGGGGCGCAGGTCACGACCCCT'CA TCCCCCACGGACGGGCCAGG60 TGGCCCACAGTCTCCGGGTCCGAGATCCACCCCGAAGC'CG CGGGACCCCGAGACCCTTGC120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'TC AGTTTAGGCCAAAAATCCCC180 (2) INFORMATION FOR SEQ ID N0:129:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL 50URCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 1102 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:129:

(2) INFORMATION FOR SEQ ID N0:130:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:130:

(2) INFORMATION FOR SEQ ID N0:131:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 2 Allele A* 3002 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:131:
GTGAGTGACCCCGCCCGGGGGCGCAGGTCACGACCCCT'CATCCCCCACGGACGGGCCAGG 60 TCGCCCACAGTCTCCGGGTCCGAGATCCACCCCGAAGC'CGCGGGACCCCGAGACCCTTGA 120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATTT'TCAGTTTAGGCCAAAAATTCCC 180 (2) INFORMATION FOR SEQ ID N0:132:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 241 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 2 Allele A* 3004 (xi) SEQUENCE DESCRIPTION: SEQ ID P10:132:

TCGCCCACAGTCTCCGGGTCCGAGATCCACCCCGAAGC:CGCGGGACCCCGAGACCCTTGA 120 CCCGGGAGAGGCCCAGGCGCCTTTACCCGGTTTCATT7.'TCAGTTTAGGCCAAAAATTCCC 180 (2) INFORMATION FOR SEQ ID N0:133:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 240 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 2 Allele A* B001 (xi) SEQUENCE DESCRIPTION: SEQ ID Pd0:133:
GTGAGTGACC CCGGCCCGGG CGCAGGTCAC GACCCCTC:AT CCCCTACGGA CGGGCCAGGT 60 _ CCGGGAGAGG CCCAGGCGCC TTTAGCCGGT TTCATTT"."CA GTTTAGGCCA AAAATCCCCC 180 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:134:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 602 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele consensus (xi) SEQUENCE DESCRIPTION: SEQ ID N0:134:

(2) INFORMATION FOR SEQ ID N0:135:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:135:

SUBSTITUTE SHEET ( rule 26 ) TCCCGGGTGT CCTGTCCATT CTCAAGATGG CCACATGC:GT GCTGGTGGAG TGTCCCATGA 540 (2) INFORMATION FOR SEQ ID N0:136:
(i} SEQUENCE CHARACTERISTICS:
(A) LENGTH: 579 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Ini:ron 3 Allele A* 0301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:136:

AGAGGAATCCTCCTGGGTTCCAGATCCTGTACCAGAGi~GTGACTCTGAGGTTCCGCCCTG180 CTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAGGi~GTGACGGGAAGACGATCCCTCG240 TCAGGCCTTGTTCTCTGCTTCACACTCAATGTGTGTGt3GGGTCTGAGTCCAGCACTTCTG360 AGTCCCTCAGCCTCCACTCAGGTCAGGACCAGAAGTC(iCTGTTCCCTTCTCAGGGAATAG420 (2) INFORMATION FOR SEQ ID N0:1:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii} HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Inl.ron 3 Allele A* 1101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:137:

AGAGGAATCC TCCTGGGTTT CCAGATCCTG TACCAGA(3AG TGACTCTGAG GTTCCGCCCT 180 GAATACTGAT GAGTGGTTCC CTTTGACACC GGCAGCA(~CC TTGGGCCCGT GACTTTTCCT 300 CTCAGGCCTT GTTCTCTGCT TCACACTCAA TGTGTGTt3GG GGTCTGAGTC CAGCACTTCT 360 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:138:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (Cj STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 1102 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:138:

(2) INFORMATION FOR SEQ ID N0:139:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:139:

SUBSTITUTE SHEET ( rule 26 ) GAATACTGATGAGTGGTTCCCTTTGACACCGGCAGCAC~CCTTGGGCCCGTGACTTTTCCT 300 GAAGATTATCCCAGGTGCCTGTGTCCAGGCTGGTGTC'CGGGTTCTGTGCTCTCTTCCCCA 480 (2) INFORMATION FOR SEQ ID N0:140:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 3002 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:140:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC:ACCCTCCCTCTGGTCCTGAGGG 220 GCTCTCTGAGCTCAATTAAGGGATAAAATCTCTGAAGC:AGTGACGGGAAGACGATCCCTC 240 GAATACTGATGAGTGGTTCCCTTTGACACCGGCAGCACiCCTTGGGCCCGTGACTTTTCCT 300 CTCAGGCCTTGTTCTCTGCTTCACACTC:AATGTGTGTGGGGGTCTGAGTCCAGCACTTCT 360 GAGTCCCTCAGCCTCCACTCAGGTCAGGACCAGAAGTC'.GCTGTTCCCTTCTCAGGGAATA 420 TCCCGGGTGTCCTGTCCATTCTCAAGATGGCCACATGC'.GTGCTGGTGGAGTGTCCCATGA 540 CAGATGCAAAATGCCTGAATTTTCTGACTCTTCCCGTC'.AG 580 (2) INFORMATION FOR SEQ ID N0:19:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int.ron 3 Allele A* 3004 (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:141:
GTACCAGGGG CCACGGGGCG CCTTCCTGAT CGCCTGT1?,GA TCTCCCGGGC TGGCCTCCCA 60 CAAGGAGGGG AGACAATTGG GACCAACACT AGAATATC'AC CCTCCCTCTG GTCCTGAGGG 120 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:142:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:142:

(2) INFORMATION FOR SEQ ID N0:143:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0202 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:143:
SUBSTITUTE SHEET ( rule 26 ) GTACCAGGGGCCACGGGGCGCCTCCCTGATCGCCTGTA.GATCTCCCGGGCTGGCCTCCCA 60 CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC'GCCCTCCCTCTGGTCCTGAGGG 120 GAATACTGATGAGTGGTTCCCTTTGACACACACAGGCP.GCAGCCTTGGGCCCGTGACTTT 300 TTCTGAGTCCTTCAGCCTCCACTCAGGTCAGGACCAGF.AGTCGCTGTTCCCTCTTCAGGG 420 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTCC'.ATTCTCAAGATAGCCACATGTG 540 (2) INFORMATION FOR SEQ ID N0:194:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 0203 (xi) SEQUENCE DESCRIPTION: SEQ ID Df0:144:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC.'GCCCTCCCTCTGGTCCTGAGGG 120 AGAGGAATCCTCCTGGGTTTCCAGATCCTGTACCAGAC:AGTGACTCTGAGGTTCCGCCCT 180 GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAGCiAATGACGGGAAGACGATCCCTC 240 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTCC'ATTCTCAAGATAGCCACATGTG 540 (2) INFORMATION FOR SEQ ID N0:19:5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 0204 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:145:

(2) INFORMATION FOR SEQ ID N0:146:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0205 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:146:

(2) INFORMATION FOR SEQ ID N0:147:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no SUBSTITUTE SHEET ( rule 26 ) (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0206 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:147:

(2) INFORMATION FOR SEQ ID N0:148:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0207 (xi) SEQUENCE DESCRIPTION: SEQ ID :N0:148:
GTACCAGGGGCCACGGGGCGCCTCCCTGATCGCCTGT.AGATCTCCCGGGCTGGCCTCCCA60 GAATACTGATGAGTGGTTCCCTTTGACACACACAGGC.AGCAGCCTTGGGCCCGTGACTTT300 TTCTGAGTCCTTCAGCCTCCACTCAGGTCAGGACCAG.AAGTCGCTGTTCCCTCTTCAGGG420 ACTAGAATTTTCCACGGAATAGGAGATTATCCCAGGT~uCCTGTGTCCAGGCTGGTGTCTG480 TGCTGGAGGAGTGTCCCATGACAGATGCAAAATGCCT~3AATGATCTGACTCTTCCTGACA600 (2) INFORMATION FOR SEQ ID N0:149:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear SUBSTITUTE SHEET ( rule 26 ) (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0208 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:149:

(2) INFORMATION FOR SEQ ID N0:150:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid {C) STRANDEDNESS: single (D) TOPOLOGY: linear {ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0209 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:150:

G

(2) INFORMATION FOR SEQ ID N0:151:
(i) SEQUENCE CHARACTERISTICS:
SUBSTITUTE SHEET ( rule 26 ) (A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In~~ron 3 Allele A* 0210 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:151:

GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAGL;AATGACGGGAAGACGATCCCTC240 GAATACTGATGAGTGGTTCCCTTTGACACACACAGGC~4GCAGCCTTGGGCCCGTGACTTT300 TTCTGAGTCCTTCAGCCTCCACTCAGGTCAGGACCAG~4AGTCGCTGTTCCCTCTTCAGGG420 TGCTGGAGGAGTGTCCCATGACAGATGCAAAATGCCT(3AATGATCTGACTCTTCCTGACA600 (2) INFORMATION FOR SEQ ID NO:I52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In1_ron 3 Allele A* 0211 (xi) SEQUENCE DESCRIPTION: SEQ ID lJ0:152:

AGAGGAATCCTCCTGGGTTTCCAGATCCTGTACCAGA(3AGTGACTCTGAGGTTCCGCCCT180 GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAG(3AATGACGGGAAGACGATCCCTC240 GAATACTGATGAGTGGTTCCCTTTGACACACACAGGCAGCAGCCTTGGGCCCGTGACTTfi300 TCCTCTCAGGCCTTGTTCTCTGCTTCACACTCAATGT(3TGTGGGGGTCTGAGTCCAGCAC360 TTCTGAGTCCTTCAGCCTCCACTCAGGTCAGGACCAGi~.P.GTCGCTGTTCCCTCTTCAGGG420 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTC(=ATTCTCAAGATAGCCACATGTG540 TGCTGGAGGAGTGTCCCATGACAGATGCAAAATGCCT(3AATGATCTGACTCTTCCTGACA600 SUBSTITUTE SHEET ( ruie 26 ) (2) INFORMATION FOR SEQ ID N0:153:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0215 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:153:

(2) INFORMATION FOR SEQ ID N0:154:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 0217 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:154:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATCGCCCTCCCTCTGGTCCTG,AGGG120 SUBSTITUTE SHEET ( rule 26 ) GGTTCTGTGC TCCCTTCCCC ATCCCAGGTG TCCTGTCC:AT TCTCAAGATA GCCACATGTG 540 TGCTGGAGGA GTGTCCCATG ACAGATGCAA AATGCCTG:AA TGATCTGACT CTTCCTGACA 600 (2) INFORMATION FOR SEQ ID N0:15.5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid _ (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 6801 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:155:

(2) INFORMATION FOR SEQ ID N0:156:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A Intron 3 Allele A* 6802 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:156:
GTACCAGGGG CCACGGGGCG CCTCCCTGAT CGCCTGTA.GA TCTCCCGGGC TGGCCTCCCA 60 SUBSTITUTE SHEET ( rule 26 ) {2) INFORMATION FOR SEQ ID N0:157:
(i) SEQUENCE CHARACTERISTICS:
{A) LENGTH: 601 base pairs (B) TYPE: nucleic acid {C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 6901 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:157:

(2) INFORMATION FOR SEQ ID N0:158:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 2301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:158:
SUBSTITUTE SHEET ( rule 26 ) CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC:GCCCTCCCTCTGGTCCTGAGGG 120 AGAGGAATCCTCCTGGGTTTCCAGATCCTGTACCAGACiAGTGACTCTGAGGTTCCGCCCT 180 GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGACGCiAATGACGGAAAGACGATCCCTC 240 GAGTCCCTCAGCCTCCACTCAGGTCAGGACCAGAAGTC:GCTGTTCCCTCCTCAGGGAATA 420 GAAGATTATCCCAGGTGCCTGTGTCCAGGCTGGTGTC7:GGGTTCTGTGCTCTCTTCCCCA 480 TCCCGGGTGTCCTGTCCATTCTCAAGATGGCCACATGC:ATGCTGGTGGAGTGTCCCATGA 540 CAGATGCAAAATGCCTGAATTTTCTGACTCTTCCCGTC:AG 580 {2) INFORMATION FOR SEQ ID N0:1_-°i9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear {ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no {v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A Int:ron 3 Allele A* 2402 (xi) SEQUENCE DESCRIPTION: SEQ ID D10:159:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC:GCCCTCCCTCTGGTCCTGAGGG 120 GAATACTGATGACTGGTTCCCTTTGACACCGGCAGCAC~CCTTGGGACCGTGACTTTTCCT 300 GAGTCCCTCAGCCTCCACTCAGGTCAGGACCAGAAGTC:GCTGTTCCCTCCTCAGGGAATA 420 GAAGATTATCCCAGGTGCCTGTGTCCAGGCTGGTGTC7.'GGGTTCTGTGCTCTCTTCCCCA 480 TCCCGGGTGTCCTGTCCATTCTCAAGATGGCCACATGC:ATGCTGGTGGAGTGTCCCATGA 540 CAGATGCAAAATGCCTGAATTTTCTGACTCTTCCCGTC:AG 580 (2) INFORMATION FOR SEQ ID NO:lEiO:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 2403 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:160:

T

{2) INFORMATION FOR SEQ ID N0:161:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single {D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 2404 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:161:

(2) INFORMATION FOR SEQ ID N0:162:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 2407 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:162:

GAAGATTATCCCAGGTGCCTGTGTCCAGGCTGGTGTC'rGGGTTCTGTGCTCTCTTCCCCA 480 (2) INFORMATION FOR SEQ ID N0:1~53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 580 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A In~tron 3 Allele A* 2407 (xi) SEQUENCE DESCRIPTION: SEQ ID 160:163:

GAAGATTATCCCAGGTGCCTGTGTCCAGGCTGGTGTC'rGGGTTCTGTGCTCTCTTCCCCA480 (2) INFORMATION FOR SEQ ID N0:164:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (vl ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
SUBSTITUTE SHEET ( rule 26 ) (A) NAME/KEY: HLA-A Intron 3 Allele A* 2501 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:164:

(2) INFORMATION FOR SEQ ID N0:165:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 2601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:165:

G

(2) INFORMATION FOR SEQ ID N0:166:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no SUBSTITUTE SHEET ( rule 26 ) (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:166:

GAATACTGATGAGTGGTTCCCTTTGACACACACCGGCA.GCAGCCTTGGGCCCGTGACTTT 300 (2) INFORMATION FOR SEQ ID N0:167:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 4301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:167:
GTACCAGGGGCCACGGGGCGCCTCCCTGATCGCCTGTA.GATCTCCCGGGCTGGCCTCCCA 60 CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC'GCCCTCCCTCTGGTCCTGAGGG 120 GAATACTGATGAGTGGTTCCCTTTGACACACACCGGCA.GCAGCCTTGGGCCCGTGACTTT 300 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTCC'ATTCTCAAGATAGCCACATGTG 540 (2) INFORMATION FOR SEQ ID N0:168:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid SUBSTITUTE SHEIET ( rule 26 ) (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 6601 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:168:

(2) INFORMATION FOR SEQ ID N0:169:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 6602 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:169:

G

(2) INFORMATION FOR SEQ ID N0:170:
SUBSTITUTE SHEET ( rule 26 ) (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 6603 (xi) SEQUENCE DESCRIPTION: SEQ ID rI0:170:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC:GCCCTCCCTCTGGTCCTGAGGG 120 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTCC:ATTCTCAAGATAGCCACATGTG 540 (2) INFORMATION FOR SEQ ID N0:171:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Int:ron 3 Allele A* 2901 (xi) SEQUENCE DESCRIPTION: SEQ ID r10:171:

CAAGGAGGGGAGACAATTGGGACCAACACTAGAATATC:GCCCTCC:CTCTGGTCCTGAGGG 120 AGAGGAATCCTCCTGGGTTTCCAGATCCTGTACCAGAC=AGTGACTCTGAGGTTCCGCCCT 180 GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAGC~AATGACGGGAAGACGATQCCTC 240 GGTTCTGTGCTCCCTTCCCCATCCCAGGTGTCCTGTCC:ATTCTCAAGATAGCCACATGTG 540 SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCTlCA97/00955 (2) INFORMATION FOR SEQ ID N0:172:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 2902 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:172:

(2) INFORMATION FOR SEQ ID N0:173:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3101 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:173:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:174:
(i) SEQUENCE CHARACTERISTICS:
(A} LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3201 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:174:

AGAGGAATCCTCCTGGGTTTCCAGATCCTGTACCAGAG.AGTGACTCTGAGGTTCCGCCCT180 GCTCTCTGAGCACAATTAAGGGATAAAATCTCTGAAGG.AATGACGGGAAGACGATCCCTC240 (2) INFORMATION FOR SEQ ID N0:175:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3301 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:175:
GTACCAGGGG CCACGGGGCG CCTCCCTGAT CGCCTGTA.GA TCTCCCGGGC TGGCCTCCCA 60 SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID NO:i76:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
{iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 3303 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:176:

G

(2) INFORMATION FOR SEQ ID N0:17?:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
{A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 7401 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID Pd0:177:

TTCTGAGTCCCTCAGCCTCCACTC.AGGTCAGGACCAGP.AGTCGCTGTTCCCTCTTCAGGG420 TGCTGGAGGAGTGTCCCATGACAGATGCAAAATGCCT(3AATGTTCTGACTCTTCCTGACA600 (2) INFORMATION FOR SEQ ID NO:1'78:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: HLA-A In~tron 3 Allele A* 7402 (xi) SEQUENCE DESCRIPTION: SEQ ID 7V0:178:

TTCTGAGTCCCTCAGCCTCCACTCAGGTCAGGACCAG~~1AGTCGCTGTTCCCTCTTCAGGG 420 (2) INFORMATION FOR SEQ ID NO:1'79:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHEET ( rule 26 ) ( i.x ) FEATURE
(A) NAME/KEY: HLA-A Intron 3 Allele A* 7403 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:179:

(2) INFORMATION FOR SEQ ID N0:180:
(i) SEøUENCE CHARACTERISTICS:
(A) LENGTH: 584 base pairs (S) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: HLA-A Intron 3 Allele A* 8001 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:180:

(2) INFORMATION FOR SEQ ID N0:181:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no SUBSTITUTE SHEET ( rule 26 ) WO 98/26091 PCT/CA9'7/00955 (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Il-230m (xi) SEQUENCE DESCRIPTION: SEQ ID N0:181:

(2) INFORMATION FOR SEQ ID N0:18.2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/~KEY: I1-226 (xi) SEQUENCE DESCRIPTION: SEQ ID 240:182:

(2) INFORMATION FOR SEQ ID NO:1l33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 base pairs (By TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (vy ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I1-221m1:1 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:183:

(2) INFORMATION FOR SEQ ID N0:184:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid SUBSTITUTE SHEET ( rule 26 ) (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I1-209 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:184:

(2) INFORMATION FOR SEQ ID N0:185:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I1-214m (xi) SEQUENCE DESCRIPTION: SEQ ID N0:185:

(2) INFORMATION FOR SEQ ID N0:186:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: I1-223d (xi) SEQUENCE DESCRIPTION: SEQ ID N0:186:

(2) INFORMATION FOR SEQ ID N0:187:
SUBSTITUTE SHEET ( rule 26 ) (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1B base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGAN2SM: human (ix) FEATURE:
(A) NAME/KEY: I1-225m (xi) SEQUENCE DESCRIFTION: SEQ ID D10:187:

(2) INFORMATION FOR SEQ ID NO:1F38:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I1-237m14 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:188:

(2) INFORMATION FOR SEQ ID N0:189:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A} NAME/KEY: I1-240 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:189:
SUBSTITUTE SHEET ( rule 26 ) GGAGGAGGGT CGGGCGGA lg (2) INFORMATION FOR SEQ ID N0:190:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 5FL-243 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:190:
AGTGTCTTCG CGGTCGCTC lg (2) INFORMATION FOR SEQ ID N0:191:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 5FR-257 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:191:

(2) INFORMATION FOR SEQ ID N0:192:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 5FR-273 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID r(0:192:

(2) INFORMATION FOR SEQ ID N0:193:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: BP202 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:193:

(2) INFORMATION FOR SEQ ID NO:1!34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: BP203 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:194:

(2) INFORMATION FOR SEQ ID N0:1:95:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
SUBSTITUTE SHEET ( rule 26 ) (A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: BP142 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:195:

(2) INFORMATION FOR SEQ ID N0:196:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-236 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:196:

(2) INFORMATION FOR SEQ ID N0:197:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-239 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:197:

(2) INFORMATION FOR SEQ ID N0:198:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear SUBSTITUTE SHEET ( rule 26 ) (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/FCEY: I3-246 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:198:

(2) INFORMATION FOR SEQ ID N0:199:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-247m6 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:199:

(2) INFORMATION FOR SEQ ID N0:200:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-249 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:200:

(2) INFORMATION FOR SEQ ID N0:201:
(i) SEQUENCE CHARACTERISTICS:
SUBSTITUTE SHEET ( rule 26 ) (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-280m18 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:201:
GCGATCGTCT TCCCGTCAC lg (2) INFORMATION FOR SEQ ID N0:202:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: I3-282 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:202:

(2) INFORMATION FOR SEQ ID N0:203:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 85 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:203:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:204:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B} TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii} HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix} FEATURE:
(A) NAME/KEY: 118 (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:204:

(2) INFORMATION FOR SEQ ID N0:2C)5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 120 {xi) SEQUENCE DESCRIPTION: SEQ ID Pd0:205:

(2) INFORMATION FOR SEQ ID N0:206:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii} HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 123 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:206:

(2) INFORMATION FOR SEQ ID N0:207:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 127 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:207:

(2) INFORMATION FOR SEQ ID N0:208:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human {ix) FEATURE:
(A) NAME/KEY: 129 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:208:

(2) INFORMATION FOR SEQ ID N0:209:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHEET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: 134 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:209:

(2) INFORMATION FOR SEQ ID N0:27.0:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/ICEY: 137 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:210:

(2) INFORMATION FOR SEQ ID N0:2:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 140 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:211:

(2) INFORMATION FOR SEQ ID N0:212:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 160 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:212:

(2) INFORMATION FOR SEQ ID N0:213:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 167 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:213:
GAGCCCCGCT TCAACGCC lg (2) INFORMATION FOR SEQ ID N0:2I4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
{iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 175 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:214:

(2) INFORMATION FOR SEQ ID N0:215:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs SUBSTITUTE SHEET ( rule 26 ) (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 193 (xi) SEQUENCE DESCRIPTION: SEQ ID PJ0:215:
GCCGGAGTAT TGGGACCG lg (2) INFORMATION FOR SEQ ID N0:2:L6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 202 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:216:

(2) INFORMATION FOR SEQ ID N0:217:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1B base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 9g (xi) SEQUENCE DESCRIPTION: SEQ ZD a~0:217:
GCAGGGTCCC CAGGTCCA lg SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:218:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 115 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:218:

(2) INFORMATION FOR SEQ ID N0:219:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 116 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:219:

(2) INFORMATION FOR SEQ ID N0:220:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 117 SUBSTITUTE SHEET ( rule 26 ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:220:

(2) INFORMATION FOR SEQ ID N0:221:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomi~~ DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 126 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:221:

(2) INFORMATION FOR SEQ ID N0:222:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 133 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:222:
GGAGCGCGAT CCGCAGGC lg (2) INFORMATION FOR SEQ ID N0:223:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SI-II:ET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: 135 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:223:

(2) INFORMATION FOR SEQ ID N0:224:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 136 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:224:

(2) INFORMATION FOR SEQ ID N0:225:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 138 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:225:

(2) INFORMATION FOR SEQ ID N0:226:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 142 (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:226:

(2) INFORMATION FOR SEQ ID N0:227:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 144 (xi) SEQUENCE DESCRIPTION: SEQ ID 1J0:227:

(2} INFORMATION FOR SEQ ID N0:228:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii} HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 145 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:228:
GCAGGGTCCC CAGGTTCG lg (2) INFORMATION FOR SEQ ID N0:229:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs SUBSTITUTE SHEET ( rule 26 ) (B) TYPE: nucleic acid (C} STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human ' (ix) FEATURE:
(A} NAME/KEY: 152 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:229:

(2) INFORMATION FOR SEQ ID N0:230:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 153 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:230:

(2) INFORMATION FOR SEQ ID N0:231:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 154 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:231:

SUBSTITUTE SHEET ( rule 26 ) (2) INFORMATION FOR SEQ ID N0:232:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 155 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:232:

(2) INFORMATION FOR SEQ ID N0:233:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 161 (xi) SEQUENCE DESCRIPTION: SEQ ID r(0:233:

(2) INFORMATION FOR SEQ ID N0:234:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 165 SUBSTITUTE SHEET ( rule 26 ) WO 98!26091 PCT/CA97/00955 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:234:

(2) INFORMATION FOR SEQ ID N0:235:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 168 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:235:

(2) INFORMATION FOR SEQ ID N0:236:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A1 ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: 180 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:236:

(2) INFORMATION FOR SEQ ID N0:237:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human SUBSTITUTE SHEET ( rule 26 ) (ix) FEATURE:
(A) NAME/KEY: Ex2 (Aw3) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:237:
GCGCCGGGAG GAGGGTC 1~
(2) INFORMATION FOR SEQ ID N0:238:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex2 (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:238:

(2) INFORMATION FOR SEQ ID N0:2_t9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex3 (xi) SEQUENCE DESCRIPTION: SEQ ID 1V0:239:

(2) INFORMATION FOR SEQ ID N0:240:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
SUBSTITUTE SHEET ( rule 26 ) (iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
{A) ORGANISM: human (ix) FEATURE:
{A) NAME/KEY: Ex3 (Aw6) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:240:

(2) INFORMATION FOR SEQ ID N0:241:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL. SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex2 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:241:

(2) INFORMATION FOR SEQ ID N0:242:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex3 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:242:
GGGCGGGGCG GGGCTCGGG ~ 19 (2) INFORMATION FOR SEQ ID N0:243:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 base pairs SUBSTITUTE SHEET ( rule 26 ) (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE '.CYPE: genomic :DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex2 (ABCwl) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:243:

(2) INFORMATION FOR SEQ ID N0:244:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
(A) ORGANISM: human (ix) FEATURE:
(A) NAME/KEY: Ex3 (ABCw2) (xi) SEQUENCE DESCRIPTION: SEQ ID rf0:244:

(2) INFORMATION FOR SEQ ID N0:295:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: genomic DNA
(iii) HYPOTHETICAL: no (iv) ANTI-SENSE: no (v) ORIGINAL SOURCE:
iA) ORGANISM: human (ix) FEATURE:
~ (A) NAME/KEY: 119 (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID r10:245:

SUBSTITUTE SHEET ( rule 26 )

Claims (26)

WHAT IS CLAIMED IS:
1. A method of determining the HLA Class I group type of a subject comprising the following steps:
(i) combining a group-specific untranslated region primer pair with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur; and (ii) determining whether a nucleic acid product is produced by the amplification;
wherein the ability of the primer pair to produce a nucleic acid product is associated with a particular HLA group type.
2. The method of claim 1, wherein the HLA Class I group to be determined is part of the HLA-A locus.
3. The method of claim 1, further comprising the step of (iii) determining the nucleic acid sequence of the nucleic acid product of step (ii).
4. The method of claim 1, wherein the primer pair comprises one or more oligonucleotide primers selected from the group consisting of I1-210m (SEQ ID
NO:35), I1-230m (SEQ ID NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID
NO:183), I1-209 (SEQ ID NO:184), I1-214m (SEQ ID NO:185), I1-223d (SEQ ID
NO:186), I1-225m (SEQ ID NO:187), I1-237m14 (SEQ ID NO:188), I1-240 (SEQ
ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ ID NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ
ID NO:198), I3-247m6 (SEQ ID NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ ID NO:202).
5. The method of claim 1, wherein the primer pair is selected from the group of pairs consisting of I1-230m (SEQ ID NO:181) and BP142 (SEQ ID
NO:195); 5'FR-257 (SEQ ID NO:191) and I3-247m6 (SEQ ID NO:199); I1-230m (SEQ ID NO:181) and I3-247m6 (SEQ ID NO:199); I1-226 (SEQ ID NO:182) and I3-249 (SEQ ID NO:200); I1-221m11 (SEQ ID NO:183) and I3-280m18 (SEQ ID

NO:201); 5'FL-243 (SEQ ID NO:190) and I3-249 (SEQ ID NO:200); I1-214m (SEQ
ID NO:185) and I3-249 (SEQ ID NO:200); I1-210m (SEQ ID NO:35) and I3-236 (SEQ ID NO:196); I1-210m (SEQ ID NO:35) and I3-249 (SEQ ID NO:200);
5'FR-273 (SEQ 117 NO:192) and I3-249 (SEQ ID NO:200); I1-223d (SEQ ID NO:186) and I3-239 (SEQ ID NO:197); I1-223d (SEQ ID NO:186) and I3-249 (SEQ ID NO:200);
I1-240 (SEQ ID NO:189) and I3-249 (SEQ ID NO:200); I1-237m14 (SEQ ID
NO:188) and I3-249 (SEQ ID NO:200); I1-225m (SEQ ID NO:187) and I3-249 (SEQ
ID NO:200); BP202 (SEQ ID NO:193) and I3-249 (SEQ ID NO:200) and BP203 (SEQ ID NO:194) and I3-282 (SEQ ID NO:202).
6. A method of determining the HLA Class I allele type of a subject comprising the following steps:
(i) combining a group-specific exon region primer pair with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur;
(ii) determining whether a first nucleic acid product is produced by the amplification wherein the ability of the primer pair to produce a nucleic acid product is associated with a particular HLA group type, and thereby identifying the group type;
(iii) combining a group-specific untranslated region primer pair corresponding to the group type of the subject, as determined by steps (i)-(ii), with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur and a second nucleic acid product is produced; and (iv) determining the nucleic acid sequence of the second nucleic acid product collected in step (iii).
7. The method of claim 6, wherein the HLA Class I allele to be determined is part of the HLA-A locus.
8. The method of claim 6, wherein the group-specific exon region primer pair used in step (i) comprises one or more oligonucleotide primers selected from the group consisting of primers 85 (SEQ ID NO:203), 118 (SEQ ID NO:204), 120 (SEQ ID NO:205), 123 (SEQ ID NO:206), 127 (SEQ ID NO:207), 129 (SEQ ID
NO:208), 134 (SEQ ID NO:209), 137 (SEQ ID NO:210), 140 (SEQ ID NO:211), 160 (SEQ ID NO:212), 167 (SEQ ID NO:213), 175 (SEQ ID NO:214), 193 (SEQ ID
NO:215), 202 (SEQ ID NO:216), 98 (SEQ ID NO:217), 115 (SEQ ID NO:218), 116 (SEQ ID NO:219), 117 (SEQ ID NO:220), 119 (SEQ ID NO:245), 126 (SEQ ID
NO:221), 133 (SEQ ID NO:222), 135 (SEQ ID NO:223), 136 (SEQ ID NO:224), 138 (SEQ ID NO:225), 142 (SEQ ID NO:226), 144 (SEQ ID NO:227), 145 (SEQ ID
NO:228), 152 (SEQ ID NO:229), 153 (SEQ ID NO:230), 154 (SEQ ID NO:231), 155 (SEQ ID NO:232), 161 (SEQ ID NO:233), 165 (SEQ ID NO:234), 168 (SEQ ID
NO:235) and 180 (SEQ ID NO:236).
9. The method of claim 6, wherein the group-specific exon region primer pair used in step (i) is selected from the group of pairs consisting of 140 (SEQ
ID NO:211) and 142 (SEQ ID NO:226); 85 (SEQ ID NO:203) and 98 (SEQ ID
NO:217); 140 (SEQ ID NO:211) and 126 (SEQ ID NO:221); 167 (SEQ ID NO:213) and 168 (SEQ ID NO:235); 118 (SEQ ID NO:204) and 119 (SEQ ID NO:245); 129 (SEQ ID NO:208) and 115 (SEQ ID NO:218); 129 (SEQ ID NO:208) and 116 (SEQ
ID NO:219) and 117 (SEQ ID NO:220); 160 (SEQ ID NO:212) and 135 (SEQ ID
NO:223); 118 (SEQ ID NO:204) and 133 (SEQ ID NO:222); 118 (SEQ ID NO:204) and 145 (SEQ ID NO:228); 134 (SEQ ID NO:209) and 155 (SEQ ID NO:232); 134 (SEQ ID NO:209) and 136 (SEQ ID NO:224); 118 (SEQ ID NO:204) and 161 (SEQ
ID NO:233); 118 (SEQ ID NO:204) and 154 (SEQ ID NO:231); 120 (SEQ ID
NO:205) and 152 (SEQ ID NO:229); 193 (SEQ ID NO:215) and 180 (SEQ ID
NO:236); 127 (SEQ ID NO:207) and 165 (SEQ ID NO:234); 137 (SEQ ID NO:210) and 145 (SEQ ID NO:228); 175 (SEQ ID NO:214) and 115 (SEQ ID NO:218) and 116 (SEQ ID NO:219); 167 (SEQ ID NO:213) and 144 (SEQ ID NO:227); 167 (SEQ
ID NO:213) and 133 (SEQ ID NO:222); 137 (SEQ ID NO:210) and 138 (SEQ ID
NO:225); 202 (SEQ ID NO:216) and 153 (SEQ ID NO:230); and 140 (SEQ ID
NO:211) and 136 (SEQ ID NO:224).
10. The method of claim 6, wherein the group-specific untranslated region primer pair used in step (iii) comprises one or more oligonucleotide primers selected from the group consisting of I1-210m (SEQ ID NO:35), I1-230m (SEQ ID
NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), I1-209 (SEQ ID
NO:184), I1-214m (SEQ ID NO:185), I1-2234 (SEQ ID NO:186), I1-225m (SEQ
ID NO:187), I1-237m14 (SEQ ID NO:188), I1-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID NO:194), I3P142 (SEQ ID NO:195), I3-236 (SEQ
ID NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID
NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ
ID NO:202).
11. The method of claim 6, wherein the group-specific untranslated region primer pair used in step (iii) is selected from the group of oligonucleotide primer pairs consisting of I1-230m (SEQ ID NO:181) and BP142 (SEQ ID NO:195);
5'FR-257 (SEQ ID NO:191) and I3-247m6 (SEQ ID NO:199); I1-230m (SEQ ID
NO:181) and I3-247m6 (SEQ ID NO:199); I1-226 (SEQ ID NO:182) and I3-249 (SEQ ID NO:200); I1-221m11 (SEQ ID NO:183) and I3-280m18 (SEQ ID NO:201);
5'FL-243 (SEQ ID NO:190) and I3-249 (SEQ ID NO:200); I1-214m (SEQ ID
NO:185) and I3-249 (SEQ ID NO:200); I1-210m (SEQ ID NO:35) and I3-236 (SEQ
ID NO:196); I1-210m (SEQ ID NO:35) and I3-249 (SEQ ID NO:200); 5'FR-273 (SEQ ID NO:192) and I3-249 (SEQ ID NO:200); I1-223d (SEQ ID NO:186) and I3-239 (SEQ ID NO:197); I1-223d (SEQ ID NO:186) and I3-249 (SEQ ID NO:200);
I1-240 (SEQ ID NO:189) and I3-249 (SEQ ID NO:200); I1-237m14 (SEQ ID NO:188) and I3-249 (SEQ ID NO:200); I1-225m (SEQ ID NO:187) and I3-249 (SEQ ID
NO:200); BP202 (SEQ ID NO:193) and I3-249 (SEQ ID NO:200) and BP203 (SEQ
ID NO:194) and I3-282 (SEQ ID NO:202).
12. A method of determining the HLA Class I allele type of a subject comprising the following steps:
(i) combining a plurality of group-specific exon region primer pairs with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur and a first nucleic acid product is produced;
(ii) determining the size of the first nucleic acid product of the amplification;
(iii) correlating the size of the first nucleic acid product with the predicted size of a fragment associated with a particular HLA type;
(iv) combining a group-specific untranslated region primer pair corresponding to the group type of the subject, as determined by steps (i)-(iii), with a target DNA sample from the subject under conditions such that primer-based amplification of the target DNA may occur and a second nucleic acid product is produced; and (v) determining the nucleic acid sequence of the second nucleic acid product.
13. The method of claim 12, wherein the HLA Class I allele to be determined is part of the HLA-A locus.
14. The method of claim 12, wherein the plurality of group-specific exon region primer pairs used in step (i) comprises one or more oligonucleotide primers selected from the group consisting of primers 85 (SEQ ID NO:203), 118 (SEQ ID NO:204), 120 (SEQ ID NO:205), 123 (SEQ ID NO:206), 127 (SEQ ID
NO:207), 129 (SEQ ID NO:208), 134 (SEQ ID NO:209), 137 (SEQ ID NO:210), 140 (SEQ ID NO:211), 160 (SEQ ID NO:212), 167 (SEQ ID NO:213), 175 (SEQ ID
NO:214), 193 (SEQ ID NO:215), 202 (SEQ ID NO:216), 98 (SEQ ID NO:217), 115 (SEQ ID NO:218), 116 (SEQ ID NO:219), 117 (SEQ ID NO:220), 126 (SEQ ID
NO:221), 133 (SEQ ID NO:222), 135 (SEQ ID NO:223), 136 (SEQ ID NO:224), 138 (SEQ ID NO:225), 142 (SEQ ID NO:226), 144 SEQ ID NO:227), 145 (SEQ ID
NO:228), 152 (SEQ ID NO:229), 153 SEQ ID NO:230), 154 (SEQ ID NO:231), 155 (SEQ ID NO:232), 161 (SEQ ID NO:233), 165 (SEQ ID NO:234), 168 (SEQ ID
NO:235) and 180 (SEQ ID NO:236).
15. The method of claim 12, wherein the plurality of group-specific exon region primer pairs used in step (i) comprises one or more oligonucleotide primer pairs selected from the group of pairs consisting of 140 (SEQ ID
NO:211) and 142 (SEQ ID NO:226); 85 (SEQ ID NO:203) and 98 (SEQ ID NO:217); 140 (SEQ ID
NO:211) and 126 (SEQ ID NO:221); 167 (SEQ ID NO:213) and 168 (SEQ ID
NO:235); 118 (SEQ ID NO:204) and 119 (SEQ ID NO:245); 129 (SEQ ID NO:208) and 115 (SEQ ID NO:218); 129 (SEQ ID NO:208) and 116 (SEQ ID NO:219) and 117 (SEQ ID NO:220); 160 (SEQ ID NO:212) and 135 (SEQ ID NO:223); 118 (SEQ
ID NO:204) and 133 (SEQ ID NO:222); 118 (SEQ ID NO:204) and 145 (SEQ ID
NO:228); 134 (SEQ ID NO:209) and 155 (SEQ ID NO:232); 134 (SEQ ID NO:209) and 136 (SEQ ID NO:224); 118 (SEQ ID NO:204) and 161 (SEQ ID NO:233); 118 (SEQ ID NO:204) and 154 (SEQ ID NO:231); 120 (SEQ ID NO:205) and 152 (SEQ
ID NO:229); 193 (SEQ ID NO:215) and 180 (SEQ ID NO:236); 127 (SEQ ID
NO:207) and 165 (SEQ ID NO:234); 137 (SEQ ID NO:210) and 145 (SEQ ID
NO:228); 175 (SEQ ID NO:214) and 115 (SEQ ID NO:218) and 116 (SEQ ID
NO:219); 167 (SEQ ID NO:213) and 144 (SEQ ID NO:227); 167 (SEQ ID NO:213) and 133 (SEQ ID NO:222); 137 (SEQ ID NO:210) and 138 (SEQ ID NO:225); 202 (SEQ ID NO:216) and 153 (SEQ ID NO:230); and 140 (SEQ ID NO:211) and 136 (SEQ ID NO:224).
16. The method of claim 12, wherein the group-specific untranslated region primer pair used in step (iv) comprises one or more oligonucleotide primers selected from the group consisting of I1-210m (SEQ ID NO:35), I1-230m (SEQ ID
NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), I1-209 (SEQ ID
NO:184), I1-214m (SEQ ID NO:185), I1-223d (SEQ ID NO:186), I1-225m (SEQ
ID NO:187), I1-237m14 (SEQ ID NO:188), I1-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ
ID NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID
NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ

ID NO:202).
17. The method of claim 12, wherein the group-specific untranslated region primer pair used in step (iv) is selected from the group of oligonucleotide primer pairs consisting of I1-230m (SEQ ID NO:181) and BP142 (SEQ ID NO:195);
5'FR-257 (SEQ ID NO:191) and I3-247m6 (SEQ ID NO:199); I1-230m (SEQ ID
NO:181) and I3-247m6 (SEQ ID NO:199); I1-226 (SEQ ID NO:182) and I3-249 (SEQ ID NO:200); I1-221m11 (SEQ ID NO:183) and I3-280m18 (SEQ ID NO:201);
5'FL-243 (SEQ ID NO:190) and I3-249 (SEQ ID NO:200); I1-214m (SEQ ID
NO:185) and I3-249 (SEQ ID NO:200); I1-210m (SEQ ID NO:35) and I3-236 (SEQ
ID NO:196); I1-210m (SEQ ID NO:35) and I3-249 (SEQ ID NO:200); 5'FR-273 (SEQ ID NO:192) and I3-249 (SEQ ID NO:200); I1-223d (SEQ ID NO:186) and I3-239 (SEQ ID NO:197); I1-223d (SEQ ID NO:186) and I3-249 (SEQ ID NO:200);
I1-240 (SEQ ID NO:189) and I3-249 (SEQ ID NO:200); I1-237m14 (SEQ ID NO:188) and I3-249 (SEQ ID NO:200); I1-225m (SEQ ID NO:187) and I3-249 (SEQ ID
NO:200); BP202 (SEQ ID NO:193) and I3-249 (SEQ ID NO:200) and BP203 (SEQ
ID NO:194) and I3-282 (SEQ ID NO:202).
18. A composition comprising a plurality of oligonucleotide primer pairs comprising one or more primers selected from the group consisting of I1-210m (SEQ ID NO:35), I1-230m (SEQ ID NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), I1-209 (SEQ ID NO:184), I1-214m (SEQ ID NO:185), I1-223d (SEQ ID NO:186), I1-225m (SEQ ID NO:187), I1-237m14 (SEQ ID NO:188), I1-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ ID NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ ID NO:202).
19. A composition comprising an oligonucleotide primer selected from the group consisting of I1-210m (SEQ ID NO:35), I1-230m (SEQ ID NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), I1-209 (SEQ ID NO:184), 214m (SEQ ID NO:185), I1-223d (SEQ ID NO:186), I1-225m (SEQ ID NO:187), I1-237m14 (SEQ ID NO:188), I1-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID
NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID
NO:193), BP203 (SEQ ID NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ ID
NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID
NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ
ID NO:202).
20. A composition comprising an oligonucleotide primer pair selected from the group consisting of I1-230m (SEQ ID NO:181) and BP142 (SEQ ID
NO:195); 5'FR-257 (SEQ ID NO:191) and I3-247m6 (SEQ ID NO:199); I1-230m (SEQ ID NO:181) and I3-247m6 (SEQ ID NO:199); I1-226 (SEQ ID NO:182) and I3-249 (SEQ ID NO:200); I1-221m11 (SEQ ID NO:183) and I3-280m18 (SEQ ID
NO:201); 5'FL-243 (SEQ ID NO:190) and I3-249 (SEQ ID NO:200); I1-214m (SEQ
ID NO:185) and I3-249 (SEQ ID NO:200); I1-210m (SEQ ID NO:35) and I3-236 (SEQ ID NO:196); I1-210m (SEQ ID NO:35) and I3-249 (SEQ ID NO:200);
5'FR-273 (SEQ ID NO:192) and I3-249 (SEQ ID NO:200); I1-223d (SEQ ID NO:186) and I3-239 (SEQ ID NO:197); I1-223d (SEQ ID NO:186) and I3-249 (SEQ ID NO:200);
I1-240 (SEQ ID NO:189) and I3-249 (SEQ ID NO:200); I1-237m14 (SEQ ID
NO:188) and I3-249 (SEQ ID NO:200); I1-225m (SEQ ID NO:187) and I3-249 (SEQ
ID NO:200); BP202 (SEQ ID NO:193) and I3-249 (SEQ ID NO:200) and BP203 (SEQ ID NO:194) and I3-282 (SEQ ID NO:202).
21. A kit comprising:
(a) a plurality of oligonucleotide; group-specific untranslated region primer pairs comprising one or more primers selected from the group consisting of I1-210m (SEQ ID NO:35), II-234m (SEQ ID NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), II-209 (SEQ ID NO:184), I1-214m (SEQ ID NO:185), I1-223d (SEQ ID NO:186), I1-225m (SEQ ID NO:187), I1-237m14 (SEQ ID
NO:188), I1-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID
NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID

NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ ID NO:196), I3-239 (SEQ ID
NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID NO:199), I3-249 (SEQ ID
NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ ID NO:202); and (b) an enzyme for nucleotide chain extension.
22. A kit comprising:
(a) an oligonucleotide group-specific untranslated region primer selected from the group consisting of I1-210m (SEQ ID NO:35), I1-230m (SEQ ID
NO:181), I1-226 (SEQ ID NO:182), I1-221m11 (SEQ ID NO:183), I1-209 (SEQ ID
NO:184), I1-214m (SEQ ID NO:185), I1-223d (SEQ ID NO:186), I1-225m (SEQ
ID NO:187), I1-237m14 (SEQ ID NO:188), II-240 (SEQ ID NO:189), 5'FL-243 (SEQ ID NO:190), 5'FR-257 (SEQ ID NO:191), 5'FR-273 (SEQ ID NO:192), BP202 (SEQ ID NO:193), BP203 (SEQ ID NO:194), BP142 (SEQ ID NO:195), I3-236 (SEQ
ID NO:196), I3-239 (SEQ ID NO:197), I3-246 (SEQ ID NO:198), I3-247m6 (SEQ ID
NO:199), I3-249 (SEQ ID NO:200), I3-280m18 (SEQ ID NO:201) and I3-282 (SEQ
ID NO:202); and (b) an enzyme for nucleotide chain extension.
23. The kit of claim 22, further comprising:
(c) a cocktail comprising group-specific exon region primers comprising one or more primer selected from the group consisting of 85 (SEQ ID
NO:203), 118 (SEQ ID NO:204), 120 (SEQ ID NO:205), 123 (SEQ ID NO:206), 127 (SEQ ID NO:207), 129 (SEQ ID NO:208), 134 (SEQ ID NO:209), 137 (SEQ ID
NO:210), 140 (SEQ ID NO:211), 160 (SEQ ID NO:212), 167 (SEQ ID NO:213), 175 (SEQ ID NO:214), 193 (SEQ ID NO:215), 202 (SEQ ID NO:216), 98 (SEQ ID
NO:217), 115 (SEQ ID NO:218), 116 (SEQ ID NO:219), 117 (SEQ ID NO:220), 126 (SEQ ID NO:221), 133 (SEQ ID NO:222), 135 (SEQ ID NO:223), 136 (SEQ ID
NO:224), 138 (SEQ ID NO:225), 142 (SEQ ID NO:226), 144 SEQ ID NO:227), 145 (SEQ ID NO:228), 152 (SEQ ID NO:229), 153 SEQ ID NO:230), 154 (SEQ ID
NO:231), 155 (SEQ ID NO:232), 161 (SEQ ID NO:233), 165 (SEQ ID NO:234), 168 (SEQ ID NO:235) and 180 (SEQ ID NO:236).
24. A kit comprising:
(a) an oligonucleotide primer pair selected from the group consisting of I1-230m (SEQ ID NO:181) and BP142 (SEQ ID NO:195); 5'FR-257 (SEQ ID
NO:191) and I3-247m6 (SEQ ID NO:199); I1-230m (SEQ ID NO:181) and I3-247m6 (SEQ ID NO:199); I1-226 (SEQ ID NO:182) and I3-249 (SEQ ID NO:200);
I1-221m11 (SEQ ID NO:183) and I3-280m18 (SEQ ID NO:201); 5'FL-243 (SEQ ID
NO:190) and I3-249 (SEQ ID NO:200); I1-214m (SEQ ID NO:185) and I3-249 (SEQ
ID NO:200); I1-210m (SEQ ID NO:35) and I3-236 (SEQ ID NO:196); I1-210m (SEQ
ID NO:35) and I3-249 (SEQ ID NO:200); 5'FR-273 (SEQ ID NO:192) and I3-249 (SEQ ID NO:200); I1-223d (SEQ ID NO:186) and I3-239 (SEQ ID NO:197); I1-223d (SEQ ID NO:186) and I3-249 (SEQ ID NO:200); I1-240 (SEQ ID NO:189) and I3-249 (SEQ ID NO:200); I1-237m14 (SEQ ID NO:188) and I3-249 (SEQ ID NO:200);
I1-225m (SEQ ID NO:187) and I3-249 (SEQ ID NO:200); BP202 (SEQ ID NO:193) and I3-249 (SEQ ID NO:200) and BP203 (SEQ ID NO:194) and I3-282 (SEQ ID
NO:202); and (b) an enzyme for nucleotide chain extension..
25. The kit of claim 24, further comprising:
(c) a cocktail comprising group-specific exon region primers comprising one or more primer selected from the group consisting of 85 (SEQ ID
NO:203), 118 (SEQ ID NO:204), 120 (SEQ ID NO:205), 123 (SEQ ID NO:206), 127 (SEQ ID NO:207), 129 (SEQ ID NO:208), 134 (SEQ ID NO:209), 137 (SEQ ID
NO:210), 140 (SEQ ID NO:211), 160 (SEQ ID NO:212), 167 (SEQ ID NO:213), 175 (SEQ ID NO:214), 193 (SEQ ID NO:215), 202 (SEQ ID NO:216), 98 (SEQ ID
NO:217), 115 (SEQ ID NO:218), 116 (SEQ ID NO:219), 117 (SEQ ID NO:220), 126 (SEQ ID NO:221), 133 (SEQ ID NO:222), 135 (SEQ ID NO:223), 136 (SEQ ID
NO:224), 138 (SEQ ID NO:225), 142 (SEQ ID NO:226), 144 SEQ ID NO:227), 145 (SEQ ID NO:228), 152 (SEQ ID NO:229), 153 SEQ ID NO:230), 154 (SEQ ID
NO:231), 155 (SEQ ID NO:232), 161 (SEQ ID NO:233), 165 (SEQ ID NO:234), 168 (SEQ ID NO:235) and 180 (SEQ ID NO:236).
(b) an enzyme for nucleotide chain extension..
26. The kit of claim 25, further comprising:
(d) a sequencing primer selected from the group consisting of 5'-EX2(Aw3) 5'GCG GCG GGA GGA GGG TC 3'(SEQ ID NO:237), 3'-Ex25'ATC
TCG GAC CCG GAG ACT 3'(SEQ ID NO:238), 5'GTT TCA TTT TCA GTT TAG
GCC A3'(SEQ ID NO:239), 3'-Ex3(Aw6)5'CGG GAG ATC TAC AGG CGA
TCA GG 3'(SEQ ID NO:241), 5'-Ex3 5'GGG CGG GGC GGG GCT CGG G 3 (SEQ ID NO:242), 3'-Ex2 (ABCwl) 5'GGT CGT GAC CT(T/C)CGC CCC 3'(SEQ
ID NO:243), and 5'-Ex3 (ABCw2) 5'CCC GGT TTC ATT TTC 3'(SEQ ID
NO:244).
CA002274737A 1996-12-12 1997-12-12 Method and kit for hla class i typing Abandoned CA2274737A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US76618996A 1996-12-12 1996-12-12
US08/766,189 1996-12-12
PCT/CA1997/000955 WO1998026091A2 (en) 1996-12-12 1997-12-12 Method and kit for hla class i typing

Publications (1)

Publication Number Publication Date
CA2274737A1 true CA2274737A1 (en) 1998-06-18

Family

ID=25075674

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002274737A Abandoned CA2274737A1 (en) 1996-12-12 1997-12-12 Method and kit for hla class i typing

Country Status (3)

Country Link
AU (1) AU5396498A (en)
CA (1) CA2274737A1 (en)
WO (1) WO1998026091A2 (en)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19715430A1 (en) * 1997-04-14 1998-11-26 Boehringer Mannheim Gmbh Method for typing alleles
JP2002510978A (en) * 1997-07-23 2002-04-09 レイクスウニヴェルシテイト レイデン Typing method of minor histocompatibility antigen HA-1
US5910413A (en) * 1997-10-10 1999-06-08 Visible Genetics, Inc. Method and kit for amplification, sequencing and typing of classical HLA class I genes
EP0953650A1 (en) 1998-04-20 1999-11-03 Innogenetics N.V. Method for typing of HLA alleles
AU6004699A (en) 1998-11-26 2000-06-13 Shionogi & Co., Ltd. Method for distinguishing hla class i allele type
MXPA01009773A (en) * 1999-04-09 2002-05-14 Innogenetics Nv Method for the amplification of hla class i alleles.
US8236771B2 (en) 2004-05-18 2012-08-07 National Institute Of Transplantation Foundation Vectors and methods for long-term immune evasion to prolong transplant viability
CA2933252A1 (en) * 2013-12-10 2015-06-18 Conexio Genomics Pty Ltd Methods and probes for identifying gene alleles

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5192659A (en) * 1989-08-25 1993-03-09 Genetype Ag Intron sequence analysis method for detection of adjacent and remote locus alleles as haplotypes
US5424184A (en) * 1991-05-08 1995-06-13 Regents Of The University Of Minnesota DNA sequence-based HLA class I typing method
EP0540997A1 (en) * 1991-11-05 1993-05-12 F. Hoffmann-La Roche Ag Methods and reagents for HLA class I DNA typing
DE4411594C1 (en) * 1994-03-30 1995-12-14 Deutsches Rheumaforschungszent Test kit for detecting HLA gene alleles by PCR amplification
US6060257A (en) * 1994-06-03 2000-05-09 Ludwig Institute For Cancer Research Tumor rejection antigens presented by HLA-B44 molecules, and uses thereof
US5997870A (en) * 1994-06-03 1999-12-07 Ludwig Institute For Cancer Research Isolated peptides which bind to HLA-B44 Molecules
GB9524381D0 (en) * 1995-11-29 1996-01-31 Anthony Nolan Bone Marrow Trus Method for identifying an unknown allele
WO1997023645A1 (en) * 1996-01-04 1997-07-03 Sloan-Kettering Institute For Cancer Research Methods and reagents for typing hla class i genes
ATE197481T1 (en) * 1996-02-20 2000-11-11 Perkin Elmer Corp METHOD AND REAGENTS FOR TYPING CLASS I HLAGENS
EP0896632B1 (en) * 1996-05-01 2001-12-12 Visible Genetics Inc. Method for detection and identification of microorganisms

Also Published As

Publication number Publication date
AU5396498A (en) 1998-07-03
WO1998026091A3 (en) 1998-09-17
WO1998026091A2 (en) 1998-06-18

Similar Documents

Publication Publication Date Title
US5834189A (en) Method for evaluation of polymorphic genetic sequences, and the use thereof in identification of HLA types
CA1242627A (en) Method for detection of polymorphic restriction sites and nucleic acid sequences
US5593830A (en) DNA sequence-based HLA class I typing method
CA2803940C (en) Application of a pcr sequencing method, based on dna barcoding technique and dna incomplete shearing strategy, in hla genotyping
EP0237362B1 (en) Process for detecting specific nucleotide variations and genetic polymorphisms present in nucleic acids and kits therefor
US6013444A (en) DNA bracketing locus compatible standards for electrophoresis
WO2001027326A2 (en) Template-dependent ligation with pna-dna chimeric probes
US5910413A (en) Method and kit for amplification, sequencing and typing of classical HLA class I genes
US6500614B1 (en) Method for identifying an unknown allele
CA2300369A1 (en) Method and kit for hla class i typing dna
CA2274737A1 (en) Method and kit for hla class i typing
EP0892069B1 (en) A method for determinig the histocompatibility locus antigen class II
WO1993018178A1 (en) DIAGNOSIS OF β-THALASSEMIA USING A MULTIPLEX AMPLIFICATION REFRACTORY MUTATION SYSTEM
US6503707B1 (en) Method for genetic typing
WO1994021818A1 (en) Process for typing hla-b using specific primers and probes sets
EP0887423A1 (en) A method for determining the Histocompatibility locus antigen class II
WO1997023645A1 (en) Methods and reagents for typing hla class i genes
Yao et al. Identification of two major HLA-B44 subtypes and a novel B44 sequence (B∗ 4404): Oligotyping and solid phase sequencing of polymerase chain reaction products
DiZinno et al. Typing of DNA derived from hairs
US20030228596A1 (en) Template-driven nucleic acid amplifications
JP2024034944A (en) Hybridization buffer composition and hybridization method
CA2009870A1 (en) Metod for determining the origin of a human dna-containing sample
US20040197775A1 (en) Intron sequence analysis method for detection of adjacent and remote locus alleles as haplotypes
Horne et al. Rapid DNA typing for class II HLA antigens: Subtyping of DRW52-associated DRB1 Alleles

Legal Events

Date Code Title Description
FZDE Discontinued