AU2006344331A1 - Modified microbial nucleic acid for use in detection and analysis of microorganisms - Google Patents

Modified microbial nucleic acid for use in detection and analysis of microorganisms Download PDF

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AU2006344331A1
AU2006344331A1 AU2006344331A AU2006344331A AU2006344331A1 AU 2006344331 A1 AU2006344331 A1 AU 2006344331A1 AU 2006344331 A AU2006344331 A AU 2006344331A AU 2006344331 A AU2006344331 A AU 2006344331A AU 2006344331 A1 AU2006344331 A1 AU 2006344331A1
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nucleic acid
seq
derivative
nucleotides
hybridizing
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AU2006344331A
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John R. Melki
Douglas Spencer Millar
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Human Genetic Signatures Pty Ltd
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Human Genetic Signatures Pty Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria

Description

WO 2007/140506 PCT/AU2006/000755 1 Modified microbial nucleic acid for use in detection and analysis of microorganisms Technical Field The invention relates to modified nucleic acid for use in detection and analysis of microorganisms. 5 Background Art A number of procedures are presently available for the detection of specific nucleic acid molecules. These procedures typically depend on sequence-dependent hybridisation between the target nucleic acid and nucleic acid probes which may range in 10 length from short oligonucleotides (20 bases or less) to sequences of many kilobases (kb). The most widely used method for amplification of specific sequences from within a population of nucleic acid sequences is that of polymerase chain reaction (PCR) (Dieffenbach, C and Dveksler, G. eds. PCR Primer: A Laboratory Manual. Cold Spring 15 Harbor Press, Plainview NY). In this amplification method, oligonucleotides, generally 20 to 30 nucleotides in length on complementary DNA strands and at either end of the region to be amplified, are used to prime DNA synthesis on denatured single-stranded DNA. Successive cycles of denaturation, primer hybridisation and DNA strand synthesis using thermostable DNA polymerases allows exponential amplification of the sequences 20 between the primers. RNA sequences can be amplified by first copying using reverse transcriptase to produce a complementary DNA (cDNA) copy. Amplified DNA fragments can be detected by a variety of means including gel electrophoresis, hybridisation with labelled probes, use of tagged primers that allow subsequent identification (eg by an enzyme linked assay), and use of fluorescently-tagged primers that give rise to a signal 25 upon hybridisation with the target DNA (eg Beacon and TaqMan systems). As well as PCR, a variety of other techniques have been developed for detection and amplification of specific nucleotide sequences. One example is the ligase chain reaction (1991, Barany, F. et al., Proc. Natl. Acad. Sci. USA 88, 189-193). Another example is isothermal amplification which was first described in 1992 30 (Walker GT, Little MC, Nadeau JG and Shank D. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. PNAS 89: 392-396 (1992) and termed Strand Displacement Amplification (SDA). Since then, a number of other isothermal amplification technologies have been described including Transcription Mediated WO 2007/140506 PCT/AU2006/000755 2 Amplification (TMA) and Nucleic Acid Sequence Based Amplification (NASBA) that use an RNA polymerase to copy RNA sequences but not corresponding genomic DNA' (Guatelli JC, Whitfield KM, Kwoh DY, Barringer KJ, Richmann DD and Gingeras TR. Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after 5 retroviral replication. PNAS 87: 1874-1878 (1990): Kievits T, van Gemen B, van Strijp D, Schukkink R, Dircks M, Adriaanse H, Malek L, Sooknanan R, Lens P. NASBA isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection. J Virol Methods. 1991 Dec; 35(3):273-86). Other DNA-based isothermal techniques include Rolling Circle Amplification (RCA) 10 in which a DNA polymerase extends a primer directed to a circular template (Fire A and Xu SQ. Rolling replication of short circles. PNAS 92: 4641-4645 (1995), Ramification Amplification (RAM) that uses a circular probe for target detection (Zhang W, Cohenford M, Lentrichia B, Isenberg HD, Simson E, Li H, Yi J, Zhang DY. Detection of Chlamydia trachomatis by isothermal ramification amplification method: a feasibility study. J Clin 15 Microbiol. 2002 Jan; 40(1):128-32.) and more recently, Helicase-Dependent isothermal DNA amplification (HDA), that uses a helicase enzyme to unwind the DNA strands instead of heat (Vincent M, Xu Y, Kong H. Helicase-dependent isothermal DNA amplification. EMBO Rep. 2004 Aug; 5(8):795-800.) Recently, isothermal methods of DNA amplification have been described (Walker 20 GT, Little MC, Nadeau JG and Shank D. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. PNAS 89: 392-396 (1992). Traditional amplification techniques rely on continuing cycles of denaturation and renaturation of the target molecules at each cycle of the amplification reaction. Heat treatment of DNA results in a certain degree of shearing of DNA molecules, thus when DNA is limiting such 25 as in the isolation of DNA from a small number of cells from a developing blastocyst, or particularly in cases when the DNA is already in a fragmented form, such as in tissue sections, paraffin blocks and ancient DNA samples, this heating-cooling cycle could further damage the DNA and result in loss of amplification signals. Isothermal methods do not rely on the continuing denaturation of the template DNA to produce single 30 stranded molecules to serve as templates from further amplification, but on enzymatic nicking of DNA molecules by specific restriction endonucleases at a constant temperature. The technique termed Strand Displacement Amplification (SDA) relies on the ability of certain restriction enzymes to nick the unmodified strand of hemi-modified DNA 35 and the ability of a 5'-3' exonuclease-deficient polymerase to extend and displace the WO 2007/140506 PCT/AU2006/000755 3 downstream strand. Exponential amplification is then achieved by coupling sense and antisense reactions in which strand displacement from the sense reaction serves as a template for the antisense reaction (Walker GT, Little MC, Nadeau JG and Shank D. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. 5 PNAS 89: 392-396 (1992). Such techniques have been used for the successful amplification of Mycobacterium tuberculosis (Walker GT, Little MC, Nadeau JG and Shank D. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. PNAS 89: 392-396 (1992), HIV-1, Hepatitis C and HPV-16 Nuovo G. J., 2000), Chlamydia trachomatis (Spears PA, Linn P, Woodard DL and Walker GT. 10 Simultaneous Strand Displacement Amplification and Fluorescence Polarization Detection of Chlamydia trachomatis. Anal. Biochem. 247: 130-137 (1997). The use of SDA to date has depended on modified phosphorthioate nucleotides in order to produce a hemi-phosphorthioate DNA duplex that on the modified strand would be resistant to enzyme cleavage, resulting in enzymic nicking instead of digestion to 15 drive the displacement reaction. Recently, however, several "nickase" enzyme have been engineered. These enzymes do not cut DNA in the traditional manner but produce a nick on one of the DNA strands. "Nickase" enzymes include N.Alwl (Xu Y, Lunnen KD and Kong H. Engineering a nicking endonuclease N.Alwl by domain swapping. PNAS 98: 12990-12995 (2001), N.BstNB1 (Morgan RD, Calvet C, Demeter M, Agra R, Kong H. 20 Characterization of the specific DNA nicking activity of restriction endonuclease N.BstNBI. Biol Chem. 2000 Nov;381(11):1123-5.) and Mlyl (Besnier CE, Kong H. Converting Mlyl endonuclease into a nicking enzyme by changing its oligomerization state. EMBO Relp. 2001 Sep;2(9):782-6. Epub 2001 Aug 23). The use of such enzymes would thus simplify the SDA procedure. 25 In addition, SDA has been improved by the use of a combination of a heat stable restriction enzyme (Aval) and Heat stable Exo-polymerase (Bst polymerase). This combination has been shown to increase amplification efficiency of the reaction from a 108 fold amplification to 1010 fold amplification so that it is possible, using this technique, to the amplification of unique single copy molecules. The resultant amplification factor 30 using the heat stable polymerase/enzyme combination is in the order of 109 (Milla M. A., Spears P. A., Pearson R. E. and Walker G. T. Use of the Restriction Enzyme Aval and Exo-Bst Polymerase in Strand Displacement Amplification Biotechniques 1997 24:392 396).
WO 2007/140506 PCT/AU2006/000755 4 To date, all isothermal DNA amplification techniques require the initial double stranded template DNA molecule to be denatured prior to the initiation of amplification. In addition, amplification is only initiated once from each priming event. For direct detection, the target nucleic acid is most commonly separated on the 5 basis of size by gel electrophoresis and transferred to a solid support prior to hybridisation with a probe complementary to the target sequence (Southern and Northern blotting). The probe may be a natural nucleic acid or analogue such as peptide nucleic acid (PNA) or locked nucleic acid (LNA) or intercalating nucleic acid (INA). The probe may be directly labelled (eg with 32 P) or an indirect detection procedure may be 10 used. Indirect procedures usually rely on incorporation into the probe of a "tag" such as biotin or digoxigenin and the probe is then detected by means such as enzyme-linked substrate conversion or chemiluminescence. Another method for direct detection of nucleic acid that has been used widely is "sandwich" hybridisation. In this method, a capture probe is coupled to a solid support 15 and the target nucleic acid, in solution, is hybridised with the bound probe. Unbound target nucleic acid is washed away and the bound nucleic acid is detected using a second probe that hybridises to the target sequences. Detection may use direct or indirect methods as outlined above. Examples of such methods include the "branched DNA" signal detection system, an example that uses the sandwich hybridization principle 20 (1991, Urdea, M. S., et al., Nucleic Acids Symp. Ser. 24,197-200). A rapidly growing area that uses nucleic acid hybridisation for direct detection of nucleic acid sequences is that of DNA microarrays, (2002, Nature Genetics, 32, [Supplement]; 2004, Cope, L.M., et al., Bioinformatics, 20, 323-331; 2004, Kendall, S.L., et al., Trends in Microbiology, 12, 537-544). In this process, individual nucleic acid species, that may range from short 25 oligonucleotides, (typically 25-mers in the Affymetrix system), to longer oligonucleotides, (typically 60-mers in the Applied Biosystems and Agilent platforms), to even longer sequences such as cDNA clones, are fixed to a solid support in a grid pattern or photolithographically synthesized on a solid support. A tagged or labelled nucleic acid population is then hybridised with the array and the level of hybridisation to each spot in 30 the array quantified. Most commonly, radioactively- or fluorescently-labelled nucleic acids (eg cRNAs or cDNAs) are used for hybridisation, though other detection systems can be employed, such as chemiluminescence. A rapidly growing area that uses nucleic acid hybridisation for direct detection of nucleic acid sequences is that of DNA micro-arrays (Young RA Biomedical discovery 35 with DNA arrays. Cell 102: 9-15 (2000); Watson A New tools. A new breed of high tech WO 2007/140506 PCT/AU2006/000755 5 detectives. Science 289:850-854 (2000)). In this process, individual nucleic acid species, that may range from oligonucleotides to longer sequences such as complementary DNA (cDNA) clones, are fixed to a solid support in a grid pattern. A tagged or labelled nucleic acid population is then hybridised with the array and the level 5 of hybridisation with each spot in the array quantified. Most commonly, radioactively- or fluorescently-labelled nucleic acids (eg cDNAs) were used for hybridisation, though other detection systems were employed. Traditional methods for the detection of microorganisms such as bacteria, yeasts and fungi and include culture of the microorganisms on selective nutrient media then 10 classification of the microorganism based on size, shape, spore production, characters such as biochemical or enzymatic reactions and specific staining properties (such as the Gram stain) as seen under conventional light microscopy. Viral species have to be grown in specialised tissue or cells then classified based on their structure and size determined by electron microscopy. A major drawback of such techniques is that not all 15 microorganisms will grow under conventional culture or cell conditions limiting the usefulness of such approaches. With bacteria, for example, such as Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae (which all cause meningitis and amongst which N. meningitidis causes both meningitis and fulminant meningococcaemia) all three species are difficult to culture. Blood culture bottles are 20 routinely examined every day for up to seven days, and subculturing is required. H. influenzae requires special medium containing both nicotinamide adenine dinucleotide and haemin and growth on Chocolate Agar Plates. Blood cultures require trypticase soy broth or brain heart infusion and the addition of various additives such as sodium polyanetholesulphonate. For microorganisms such as Clostridium botulinum, which 25 causes severe food poisoning and floppy baby syndrome, the identification of the toxin involves injection of food extracts or culture supernatants into mice and visualization of results after 2 days. In addition, culturing of the potential microorganism on special media takes a week. Staphylococcus aureus enterotoxin (a cause of food poisoning as well as skin infections, blood infections, pneumonia, osteomyelitis, arthritis and brain 30 abscesses) is detected in minute amounts by selective absorption of the toxin via ion exchange resins or Reverse Passive Latex Agglutination using monoclonal antibodies. Its relative, S. epidermis, leads to blood infections and contaminates equipment and surfaces in hospitals and health care machines and appliances. Non-viral microorganisms can also be classified based on their metabolic 35 properties such as the production of specific amino acids or metabolites during WO 2007/140506 PCT/AU2006/000755 6 fermentation reactions on substrates such as glucose, maltose or sucrose. Alternatively, microorganisms can be typed based on their sensitivity to antibiotics. Specific antibodies to cell surface antigens or excreted proteins such as toxins are also used to identify or type microorganisms. However, all the above methods rely on the culture of the 5 microorganism prior to subsequent testing. Culture of microorganisms is expensive and time consuming and can also suffer from contamination or overgrowth by less fastidious microorganisms. The techniques are also relatively crude in that many tests must be done on the same sample in order to reach definitive diagnosis. Most microorganisms can not be readily grown in known media, and hence they fall below levels of detection 10 when a typical mixed population of different species of microorganism is present in the wild or in association with higher organisms. Other methods for the detection and identification of pathogenic microorganisms are based on the serological approach in which antibodies are produced in response to infection with the microorganism. Meningococci, for example, are classifiable on the 15 basis of the structural differences in their capsular polysaccharides. These have different antigenicities, allowing five major serogroups to be determined, (A, B, C, Y and W-135). Enzyme Linked Immunosorbent Assays (ELISA) or Radio Immuno Assay (RIA) can assess the production of such antibodies. Both these methods detect the presence of specific antibodies produced by the host animal during the course of infection. These 20 methods suffer the drawback in that it takes some time for an antibody to be produced by the host animal, thus very early infections are often missed. In addition, the use of such assays cannot reliably differentiate between past and active infection. More recently, there has been much interest in the use of molecular methods for the diagnosis of infectious disease. These methods offer sensitive and specific detection 25 of pathogenic microorganisms. Examples of such methods include the "branched DNA" signal detection system. This method is an example that uses the sandwich hybridization principle (Urdea MS et al. Branched DNA amplification multimers for the sensitive, direct detection of human HIV and hepatitis viruses. Nucleic Acids Symp Ser. 1991;(24):197-200). 30 Another method for the detection and classification of bacteria is the amplification of 16S ribosomal RNA sequences. 16S rRNA has been reported to be a suitable target for use in PCR amplification assays for the detection of bacterial species in a variety of clinical or environmental samples and has frequently been used to identify various specific microorganisms because 16S rRNA genes show species-specific 35 polymorphisms (Cloud, J. L., H. Neal, R. Rosenberry, C. Y. Turenne, M. Jama, D. R.
WO 2007/140506 PCT/AU2006/000755 7 Hillyard, and K. C. Carroll. 2002. J. Clin. Microbiol. 40:400-406). However, pure culture of bacteria are required and after PCR amplification the sample still has to be sequenced or hybridized to a micro-array type device to determine the species (Fukushima M, Kakinuma K, Hayashi H, Nagai H, Ito K, Kawaguchi R. J Clin Microbiol. 2003 Jun; 5 41(6):2605-15). Such methods are expensive, time consuming and labour intensive. Although the genomes of most microorganisms have been determined by sequence analysis, there are still problems in obtaining specific probes or primers to detect microorganisms of interest. As genomes contain four bases, often it is difficult to prepare sufficient degenerate primers or probes that will be specific for a given 10 microorganism. Another potential problem is that the rights to use some important genes or genomes of particular microorganisms are owned through patent rights. This ownership can prevent or delay competing detection assays coming to market. There is a need for new nucleic acids that can be used as markers for specific microorganisms. The present inventors have obtained modified nucleic acids for numerous 15 microorganisms that are microbial specific and can be used for detecting microorganisms. Summary of Invention In one aspect, the present invention provides a derivative or modified nucleic acid 20 for Hepatitis C virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 76 in Sequence Listing #51, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Acinetobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #1, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Bacillus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #2, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 8 In another aspect, the present invention provides a derivative or modified nucleic acid for Bacteroides sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #3, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 5 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Bartonella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #4, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 10 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Bordetella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #5, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 15 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Borrelia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #6, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 20 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Brucella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #7, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 25 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Campylobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #8, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 30 stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Chlamydia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #9, parts thereof comprising at least WO 2007/140506 PCT/AU2006/000755 9 about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Clostridium sp having a sequence selected from the group consisting of 5 SEQ ID NO: 1 to SEQ ID NO: 39 in Sequence Listing #10, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Cornebacterium sp having a sequence selected from the group consisting of 10 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #11, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Escherichia coli having a sequence selected from the group consisting of 15 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #12, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Ehrlichia sp having a sequence selected from the group consisting of 20 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #13, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Enterococcus sp having a sequence selected from the group consisting of 25 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #14, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Fusobacteriumsp having a sequence selected from the group consisting of 30 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #15, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Haemophilus sp having a sequence selected from the group consisting of WO 2007/140506 PCT/AU2006/000755 10 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #16, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 5 acid for Helicobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #17, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 10 acid for Legionella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #18, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 15 acid for Leptospira sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #19, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 20 acid for Listeria sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #20, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Mycobacterium sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #21, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Mycoplasma sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #22, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 11 In another aspect, the present invention provides a derivative or modified nucleic acid for Neisseria sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #23, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 5 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Norcadia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #24, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 10 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Pseudomonas sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #25, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 15 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Rickettsia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #26, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 20 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Salmonella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #27, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 25 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Seratia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #28, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 30 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Shigella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #29, parts thereof comprising at WO 2007/140506 PCT/AU2006/000755 12 least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Staphylococcus sp having a sequence selected from the group consisting of 5 SEQ ID NO: 1 to SEQ ID NO: 52 in Sequence Listing #30, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Streptococcus sp having a sequence selected from the group consisting of 10 SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #31, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Streptomyces having a sequence selected from the group consisting of 15 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #32, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Treponema sp having a sequence selected from the group consisting of 20 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #33, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Trophermya sp having a sequence selected from the group consisting of 25 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #34, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Plasmodium sp having a sequence selected from the group consisting of 30 SEQ ID NO: 1 to SEQ ID NO: 60 in Sequence Listing #35, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Aspergillis sp having a sequence selected from the group consisting of WO 2007/140506 PCT/AU2006/000755 13 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #36, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides. a derivative or modified nucleic 5 acid for Candida sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #37, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 10 acid for Cryptococcus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #38, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 15 acid for Paracoccidioides sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #39, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 20 acid for Rhizopus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #40, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Francisella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #41, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Vibrio sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #42, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 14 In another aspect, the present invention provides a derivative or modified nucleic acid for Yersinia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #43, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 5 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for JC polyomavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #44, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 10 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Andes virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #46, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 15 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for hepatitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #52, parts thereof comprising at 20 least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Human Immunodeficiency virus (HIV) having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 128 in Sequence Listing #53, parts 25 thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Influenza virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 162 in Sequence Listing #54, parts thereof comprising at 30 least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for BK virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #55, parts thereof comprising at least about 20 WO 2007/140506 PCT/AU2006/000755 15 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Barmah virus having a sequence selected from the group consisting of 5 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #56, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Calcivirus virus having a sequence selected from the group consisting of 10 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #57, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Colorado tick fever virus having a sequence selected from the group consisting 15 of SEQ ID NO: 1 to SEQ ID NO: 48 in Sequence Listing #58, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Foot and Mouth virus having a sequence selected from the group consisting of 20 SEQ ID NO: 1 to SEQ ID NO: 28 in Sequence Listing #59, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Hepatitis GB virus having a sequence selected from the group consisting of 25 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #60, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Henda virus having a sequence selected from the group consisting of 30 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #61, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Human adenovirus having a sequence selected from the group consisting of WO 2007/140506 PCT/AU2006/000755 16 SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #62, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 5, acid for Human astrovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #63, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 10 acid for Human bocavirus virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #64, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 15 acid for Human coronavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #65, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 20 acid for Human enterovirus virus having a sequence selected from the group'consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #66, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Human herpes virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 36 in Sequence Listing #67, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Human metapneumovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #68, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 17 In another aspect, the present invention provides a derivative or modified nucleic acid for Human parainfluenzavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #69, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 5 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Human parechovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #70, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 10 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Human rhinovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #71, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 15 under stringent conditions. In another aspect, the present'invention provides a derivative or modified nucleic acid for Human respiratory syncytial virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #72, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of 20 hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Measles virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #73, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 25 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Mumps virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #74, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 30 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Norovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #75, parts thereof comprising at WO 2007/140506 PCT/AU2006/000755 18 least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Norwalk virus having a sequence selected from the group consisting of 5 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #76, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Parvovirus Bl9having a sequence selected from the group consisting of 10 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #77, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Poliovirus having a sequence selected from the group consisting of 15 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #78, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Rabies virus having a sequence selected from the group consisting of 20 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #79, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Ross River virus having a sequence selected from the group consisting of 25 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #80, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Rotavirus having a sequence selected from the group consisting of 30 SEQ ID NO: 1 to SEQ ID NO: 124 in Sequence Listing #81, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for SARS coronavirus having a sequence selected from the group consisting of WO 2007/140506 PCT/AU2006/000755 19 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #82, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 5 acid for TT virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #83, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 10 acid for TTV minivirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #84, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 15 acid for West Nile virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #85, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 20 acid for Alpha virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #86, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Camel pox virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID: NO: 4 in Sequence Listing #87, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Cow Pox virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #88, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 20 In another aspect, the present invention provides a derivative or modified nucleic acid for Coxiella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #89, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 5 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Crimean-Congo HF having a sequence selected from the group consisting of SEQ ID NO: 1Ito SEQ ID NO: 12 in Sequence Listing #90, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 10 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Dengue virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #91, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 15 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Eastern Equine Encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #92, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of 20 hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Ebola virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #93, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 25 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Marburg virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #94, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 30 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Guanarito virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #95, parts thereof comprising at WO 2007/140506 PCT/AU2006/000755 21 least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Hanta virus having a sequence selected from the group consisting of 5 SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #96, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Hantan virus having a sequence selected from the group consisting of 10 SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #97, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Japanese encephalitis virus having a sequence selected from the group 15 consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #97, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Junin virus having a sequence selected from the group consisting of 20 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #99, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Lassa virus having a sequence selected from the group consisting of 25 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #100, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Machupo virus having a sequence selected from the group consisting of 30 SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #101, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Monkey pox virus having a sequence selected from the group consisting of WO 2007/140506 PCT/AU2006/000755 22 SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #102, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 5 acid for Murray Valley encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #103, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 10 acid for Nipah virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #104, parts thereof comprising at least about 20 nucleotides,'and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 15 acid for Rift Valley Fever virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #105, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 20 -acid for Sabia virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #106, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 25 acid for Sin Nombre virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #107, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic 30 acid for Variola major virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #108, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
WO 2007/140506 PCT/AU2006/000755 23 In another aspect, the present invention provides a derivative or modified nucleic acid for Variola minor virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #109, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 5 under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Venezuelan equine encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #110, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of 10 hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Western equine encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #111, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of 15 hybridizing thereto under stringent conditions. In another aspect, the present invention provides a derivative or modified nucleic acid for Yellow Fever virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #112, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto 20 under stringent conditions. The parts of the derivative or microbial nucleic acid can be at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60 70, 80, 90, 100, etc, or more nucleotides. In some derivative or microbial nucleic acid, the part thereof may be less than 20 such as 15, 16, 17, 18 or 19, for example. 25 Derivative microbial nucleic acid can be formed by treating microbial nucleic acid with an agent such as bisulphate that modifies cytosine to uracil. After amplification of the derivative nucleic acid, modified microbial nucleic acid is formed having substantially the bases adenine, guanine and thymine. For double stranded DNA which typically contains no methylated cytosines, the 30 treating step results in two derivative nucleic acids, each containing the bases adenine, guanine, thymine and uracil. The two derivative nucleic acids are produced from the two single strands of the double stranded DNA. The two derivative nucleic acids have substantially no cytosines but still have the same total number of bases and sequence length as the original untreated DNA molecule. Importantly, the two derivatives are not WO 2007/140506 PCT/AU2006/000755 24 complimentary to each other and form a top and a bottom strand. One or more of the strands can be used to generate a derivative nucleic acid or amplified to produce the modified nucleic acid molecule. Throughout this specification, unless the context requires otherwise, the word 5 "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. Any discussion of documents, acts, materials, devices, articles or the like which 10 has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this specification. 15 In order that the present invention may be more clearly understood, preferred embodiments will be described with reference to the following drawings and examples. Brief Description of the Drawings Figure 1 shows Hepatitis C la genome (top strand) sequence (SEQ ID NO: 77 of 20 Sequence Listing #51). Figure 2 shows Hepatitis C la genome (bottom strand) sequence (SEQ ID NO: 78 of Sequence Listing #51). Figure 3 shows Hepatitis C l a derivative genome (top strand) sequence (SEQ ID NO: 1 of Sequence Listing #51). 25 Figure 4 shows Hepatitis C la derivative genome (bottom strand) sequence (SEQ ID NO: 20 of Sequence Listing #51). Figure 5 shows Hepatitis C la modified genome (top strand) sequence (SEQ ID NO: 39 of Sequence Listing #51). Figure 6 shows Hepatitis C la modified genome (bottom strand) sequence 30 (SEQ ID NO: 58 of Sequence Listing #51).
WO 2007/140506 PCT/AU2006/000755 25 Mode(s) for Carryingq Out the Invention Definitions The term "genomic modification" as used herein means the genomic (or other) nucleic acid is modified from being comprised offour bases adenine (A), guanine (G), 5 thymine (T) and cytosine (C) to substantially containing the bases adenine (A), guanine (G), thymine (T) but still having substantially the same total number of bases. The term "derivative nucleic acid "as used herein means a nucleic acid that substantially contains the bases A, G, T and U (or some other non-A, G or T base or base-like entity) and has substantially the same total number of bases as the 10 corresponding unmodified microbial nucleic acid. Substantially all cytosines in the microbial DNA will have been converted to uracil during treatment with the agent. It will be appreciated that altered cytosines, such as by methylation, may not necessarily be converted to uracil (or some other non-A, G or T base or base-like entity). As microbial nucleic acid typically does not contain methylated cytosine (or other cytosine alterations) 15 the treated step preferably converts all cytosines. Preferably, cytosine is modified to uracil. The term "modified nucleic acid" as used herein means the resulting nucleic acid product obtained after amplifying derivative nucleic acid. Uracil in the derivative nucleic acid is then replaced as a thymine (T) during amplification of the derivative nucleic acid 20 to form the modified nucleic acid molecule. The resulting product has substantially the same number of total bases as the corresponding unmodified microbial nucleic acid but is substantially made up of a combination of three bases (A, G and T). The term "modified sequence" as used herein means the resulting nucleic acid sequence obtained after amplifying derivative nucleic acid to form a modified nucleic 25 acid. The resulting modified sequence has substantially the same number of total bases as the corresponding unmodified microbial nucleic acid sequence but is substantially made up of a combination of three bases (A, G and T). The term "non-converted sequence" as used herein means the nucleic acid sequence of the microbial nucleic acid prior to treatment. A non-converted sequence 30 typically is the sequence of the naturally occurring microbial nucleic acid. The term "modifies" as used herein means the conversion of a cytosine to another nucleotide. Preferably, the agent modifies cytosine to uracil to form a derivative nucleic acid.
WO 2007/140506 PCT/AU2006/000755 26 The term "agent that modifies cytosine" as used herein means an agent that is capable of converting cytosine to another chemical entity. Preferably, the agent modifies cytosine to uracil which is then replaced as a thymine during amplification of the derivative nucleic acid. Preferably, the agent used for modifying cytosine is sodium 5 bisulfite. Other agents that similarly modify cytosine, but not methylated cytosine can also be used in the method of the invention. Examples include, but not limited to bisulfite, acetate or citrate. Preferably, the agent is sodium bisulfite, a reagent, which in the presence of acidic aqueous conditions, modifies cytosine into uracil. Sodium bisulfite (NaHSO 3 ) reacts readily with the 5,6-double bond of cytosine to form a sulfonated 10 cytosine reaction intermediate which is susceptible to deamination, and in the presence of water gives rise to a uracil sulfite. If necessary, the sulfite group can be removed under mild alkaline conditions, resulting in the formation of uracil. Thus, potentially all cytosines will be converted to uracils. Any methylated cytosines, however, cannot be converted by the modifying reagent due to protection by methylation. It will be 15 appreciated that cytosine (or any other base) could be modified by enzymatic means to achieve a derivative nucleic acid as taught by the present invention. There are two broad generic methods by which bases in nucleic acids may be modified: chemical and enzymatic. Thus, modification for the present invention can also be carried out by naturally occurring enzymes, or by yet to be reported artificially '20 constructed or selected enzymes. Chemical treatment, such as bisulphite methodologies, can convert cytosine to uracil via appropriate chemical steps. Similarly, cytosine deaminases, for example, may carry out a conversion to form a derivative nucleic acid. The first report on cytosine deaminases to our knowledge is 1932, Schmidt, G., Z. physiol. Chem., 208, 185; (see also 1950, Wang, T.P., Sable, H.Z., Lampen, J.O., 25 J. Biol. Chem, 184, 17-28, Enzymatic deamination of cytosines nucleosides). In this early work, cytosine deaminase was not obtained free of other nucleo-deaminases, however, Wang et al. were able to purify such an activity from yeast and E. coil. Thus any enzymatic conversion of cytosine to form a derivative nucleic acid which ultimately results in the insertion of a base during the next replication at that position, that is 30 different to a cytosine, will yield a modified genome. The chemical and enzymatic conversion to yield a derivative followed by a modified genome are applicable to any nucleo-base, be it purines or pyrimidines in naturally occurring nucleic acids of microorganisms. The term "modified form of the genome or nucleic acid" as used herein means 35 that a genome or nucleic acid, whether naturally occurring or synthetic, which usually WO 2007/140506 PCT/AU2006/000755 27 contains the four common bases G, A, T and C, now consists largely of only three bases, G, A and T since most or all of the Cs in the genome have been converted to Ts by appropriate chemical modification and subsequent amplification procedures. The modified form of the genome means that relative genomic complexity is reduced from a 5 four base foundation towards a three base composition. The term 'base-like entity' as used herein means an entity that is formed by modification of cytosine. A base-like entity can be recognised by a DNA polymerase during amplification of a derivative nucleic acid and the polymerase causes A, G or T to be placed on a newly formed complementary DNA strand at the position opposite the 10 base-like entity in the derivate nucleic acid. Typically, the base-like entity is uracil that has been modified from cytosine in the corresponding untreated microbial nucleic acid. Examples of a base-like entity includes any nucleo-base, be it purine or pyrimidine. The term "relative complexity reduction" as used herein relates to probe length, namely the increase in average probe length that is required to achieve the same 15 specificity and level of hybridization of a probe to a specific locus, under a given set of molecular conditions in two genomes of the same size, where the first genome is "as is" and consists of the four bases, G, A T and C, whereas the second genome is of exactly the same length but some cytosines, (ideally all cytosines), have been converted to thymines. The locus under test is in the same location in the original unconverted as well 20 as the converted genome. On average, an 11-mer probe will have a unique location to which it will hybridize perfectly in a regular genome of 4,194,304 bases consisting of the four bases G, A, T and C, (411 equals 4,194,304). However, once such a regular genome of 4,194, 304 bases has been converted by bisulfite or other suitable means, this converted genome is now composed of only three bases and is clearly less complex. 25 However the consequence of this decrease in genomic complexity is that our previously unique 11-mer probe no longer has a unique site to which it can hybridize within the modified genome. There are now many other possible equivalent locations of 11 base sequences that have arisen de novo as a consequence of the bisulfite conversion. It will now require a 14-mer probe to find and hybridize to the original locus. Although it may 30 initially appear counter intuitive, one thus requires an increased probe length to detect the original location in what is now a modified three base genome, because more of the genome looks the same, (it has more similar sequences). Thus the reduced relative genomic complexity, (or simplicity of the three base genome), means that one has to design longer probes to find the original unique site.
WO 2007/140506 PCT/AU2006/000755 28 The term "relative genomic complexity reduction" as used herein can be measured by increased probe lengths capable of being microbe-specific as compared with unmodified DNA. This term also incorporates the type of probe sequences that are used in determining the presence of a microorganism. These probes may have non 5 conventional backbones, such as those of PNA or LNA or modified additions to a backbone such as those described in INA. Thus, a genome is considered to have reduced relative complexity, irrespective of whether the probe has additional components such as Intercalating pseudonucleotides, such as in INA. Examples include, but not limited to, DNA, RNA, locked nucleic acid (LNA), peptide nucleic acid (PNA), MNA, 10 altritol nucleic acid (ANA), hexitol nucleic acid (HNA), intercalating nucleic acid (INA), cyclohexanyl nucleic acid (CNA) and mixtures thereof and hybrids thereof, as well as phosphorous atom modifications thereof, such as but not limited to phosphorothioates, methyl phospholates, phosphoramidites, phosphorodithiates, phosphoroselenoates, phosphotriesters and phosphoboranoates. Non-naturally occurring nucleotides include, 15 but not limited to the nucleotides comprised within DNA, RNA, PNA, INA, HNA, MNA, ANA, LNA, CNA, CeNA, TNA, (2'-NH)-TNA, (3'-NH)-TNA, ca-L-Ribo-LNA, a-L-Xylo-LNA, P3-D-Xylo-LNA, -D-Ribo-LNA, [3.2.1]-LNA, Bicyclo-DNA, 6-Amino-Bicyclo-DNA, 5-epi Bicyclo-DNA, a-Bicyclo-DNA, Tricyclo-DNA, Bicyclo[4.3.0]-DNA, Bicyclo[3.2.1]-DNA, Bicyclo[4.3.0]amide-DNA, P.-D-Ribopyranosyl-NA, a-L-Lyxopyranosyl-NA, 2'-R-RNA, a-L 20 RNA or a-D-RNA, p-D-RNA. In addition non-phosphorous containing compounds may be used for linking to nucleotides such as but not limited to methyliminomethyl, formacetate, thioformacetate and linking groups comprising amides. In particular nucleic acids and nucleic acid -analogues may comprise one or more intercalator pseudonucleotides (IPN). The presence of IPN is not part of the complexity description 25 for nucleic acid molecules, nor is the backbone part of that complexity, such as in PNA. By 'INA' is meant an intercalating nucleic acid in accordance with the teaching of WO 03/051901, WO 03/052132, WO 03/052133 and WO 03/052134 (Unest A/S) incorporated herein by reference. An INA is an oligonucleotide or oligonucleotide analogue comprising one or more intercalator pseudonucleotide (IPN) molecules. 30 By 'HNA' is meant nucleic acids as for example described by Van Aetschot et al., 1995. By 'MNA' is meant nucleic acids as described by Hossain et al, 1998. 'ANA' refers to nucleic acids described by Allert et al, 1999.
WO 2007/140506 PCT/AU2006/000755 29 'LNA' may be any LNA molecule as described in WO 99/14226 (Exiqon), preferably, LNA is selected from the molecules depicted in the abstract of WO 99/14226. More preferably, LNA is a nucleic acid as described in Singh et al, 1998, Koshkin et al, 1998 or Obika et al., 1997. 5 'PNA' refers to peptide nucleic acids as for example described by Nielsen et al, 1991. 'Relative complexity reduction' as used herein, does not refer to the order in which bases occur, such as any mathematical complexity difference between a sequence that is ATATATATATATAT (SEQ ID NO: ) versus one of the same length that 10 is AAAAAAATTTTTTT (SEQ ID NO: ), nor does it refer to the original re-association data of relative genome sizes, (and inferentially, genomic complexities), introduced into the scientific literature by Waring, M. & Britten R. J.1966, Science, 154, 791-794; and Britten, R.J and Kohne D E., 1968, Science, 161, 529-540, and earlier references therein that stem from the Carnegie Institution of Washington Yearbook reports. 15 'Relative genomic complexity' as used herein refers to an unchanged position of bases in two genomes that is accessed by molecular probes (both the original and unconverted genomes have bases at invariant positions 1 to n. In the case of the 3 billion base pair haploid human genome of a particular human female, the invariant positions are defined as being from 1 to n, where n is 3,000,000,000. If in the sequence 20 1 to n, the ith base is a C in the original genome, then the ith base is a T in the converted genome. The term "genomic nucleic acid" as used herein includes microbial (prokaryote and single celled eukaryote) RNA, DNA, protein encoding nucleic acid, non-protein encoding nucleic acid, and ribosomal gene regions of prokaryotes and single celled 25 eukaryotic microorganisms. The term "microbial genome" as used herein covers chromosomal as well as extrachromosomal nucleic acids, as well as temporary residents of that genome, such a plasmids, bacteriphage and mobile elements in the broadest sense. The "genome" has a core component as exemplified by S. galactiae, as well as possibly having coding and 30 non-coding elements that vary between different isolates. The term "microbial derived DNA" as used herein includes DNA obtained directly from a microorganism or obtained indirectly by converting microbial RNA to DNA by any of the known or suitable method such as reverse transcriptase.
WO 2007/140506 PCT/AU2006/000755 30 The term "microorganism" as used herein includes phage, virus, viroid, bacterium, fungus, alga, protozoan, spirochaete, single cell organism, or any other microorganism, no matter how variously classified, such as the Kingdom Protoctista by Margulis, L., et al 990, Handbook of Protoctista, Jones and Bartlett, Publishers, Boston 5 USA, or microorganisms that are associated with humans, as defined in Harrisons Principles of Internal Medicine, 12 th Edition, edited by J D Wilson et al., McGraw Hill Inc, as well as later editions. It also includes all microorganisms described in association with human conditions defined in OMIM, Online Mendelian Inheritance in Man, www.ncbi.qov. The term "microbial-specific nucleic acid molecule" as used herein means a 10 molecule which has been determined or obtained using the method according to the present invention which has one or more sequences specific to a microorganism. The term "taxonomic level of the microorganism" as used herein includes family, genus, species, strain, type, or different populations from the same or different geographic or benthic populations. While in the case of bacteria the generally 15 recognized schema, such as; Bacteria, Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae; Neisseria is used. Different populations may be polymorphic for single nucleotide changes or variation that exists in DNA molecules that exist in an intracellular form within a microorganism (plasmids or phagemids), or polymorphic chromosomal regions of microorganism genomes such as pathogenicity islands. The fluidity of 20 microbial and viral genomes is recognized, and includes the chimeric nature of viral genomes, which can be in independent nucleic acid pieces. Hence, newly arising strains from re-assortment of genomic regions from different animals .e.g., new human influenza strains as chimeras of segments that are picked up from other mammalian or avian viral genomes. 25 The term "close sequence similarity" as used herein includes the above definition of relative sequence complexity and probe lengths as a measure. The term "hybridizing under stringent conditions" is used interchangeably with the term "capable of hybridizing under stringent conditions" herein to mean that nucleic acids may be readily identified by their ability to hybridize under stringent conditions with all or 30 parts of a modified microbial nucleic acid. By capable of hybridizing under stringent conditions it is meant that annealing of nucleic acid occurs under standard conditions, e.g., high temperature and/or low salt content, which tend to preclude hybridization of noncomplementary nucleotide sequences. An example of a stringent protocol for hybridization of nucleic acid probes to immobilised DNA (involving 0. 1xSSC, 68 0 C for 2 35 hours) is described in Maniatis, T., et al., Molecular Cloning: A Laboratory Manual, Cold WO 2007/140506 PCT/AU2006/000755 31 Springs Harbor Laboratory, 1982,.at pages 387-389, although conditions will vary depending on the application. MATERIALS and METHODS 5 Extraction of DNA In general, microbial DNA (or viral RNA) can be obtained from any suitable source. Examples include, but not limited to, cell cultures, broth cultures, environmental samples, clinical samples, bodily fluids, liquid samples, solid samples such as tissue. Microbial DNA from samples can be obtained by standard procedures. An example of a 10 suitable extraction is as follows. The sample of interest is placed in 400 pl of 7 M Guanidinium hydrochloride, 5mM EDTA, 100 mM Tris/HCI pH 6.4, 1% Triton-X-100, 50 mM Proteinase K (Sigma), 100 pg/ml yeast tRNA. The sample is thoroughly homogenised with disposable 1.5 ml pestle and left for 48 hours at 600C. After incubation the sample is subjected to five freeze/thaw cycles of dry ice for 5 15 minutes/95 0 C for 5 minutes. The sample is then vortexed and spun in a microfuge for 2 minutes to pellet the cell debris. The supernatant is removed into a clean tube, diluted to reduce the salt concentration then phenol:chloroform extracted, ethanol precipitated and resuspended in 50 pl of 10 mM Tris/0.1 mM EDTA. Specifically, the DNA extractions from Gram positive and Gram negative bacteria 20 grown on standard agar plates (with nutritional requirements specific to each species) were performed as follows. For DNA extraction from Gram Negative bacteria the protocol was as follows: a) Using a sterile toothpick bacterial colonies were scraped off the culture plate into a sterile 1.5 ml centrifuge tube. 25 b) 180 pI of Guanidinium thiocyanate extraction buffer (7M Guanidinium thiocyanate, 5 mM EDTA (pH8.0), 40 mM Tris/Hcl pH 7.6, 1% Triton-X-100) was added and the sample mixed to resuspend the bacterial colonies. c) 20 pl (20 mg/ml) Proteinase K was added and the samples were mixed well. d) Samples were incubated @ 550C for 3 hours to lyse the cells. 30 e) 200 pl of water was added to each sample and samples mixed by gentle pipetting. f) 400 pl of Phenol/Chloroform/iso-amyl alcohol (25:24:1) was added and the samples vortexed for 2 X 15 seconds. g) The samples were then spun in a microfuge at 14,000 rpm for 4 minutes. h) The aqueous phase was removed into a clean 1.5 ml centrifuge tube.
WO 2007/140506 PCT/AU2006/000755 32 i) 400 pl of Phenol/Chloroform/iso-amyl alcohol (25:24:1) was added and the samples vortexed for 2 X 15 seconds. j) The samples were then spun in a microfuge at 14,000rpm for 4 minutes. k) The aqueous phase was removed into a clean 1.5 ml centrifuge tube. 5 I) 800 pl of 100% ethanol was added to each sample, the sample vortexed briefly then left at -20 0 c for 1 hour. m) The samples were spun in a microfuge at 14,000 rpm for 4 minutes at 40C. n) The DNA pellets were washed with 500 pl of 70% ethanol. o) The samples were spun in a microfuge at 14,000rpm for 5 minutes at 4 0 C, the 10 ethanol was discarded and the pellets were air dried for 5 minutes. p) Finally the DNA was resuspended in 100 pl of 10 mM Tris/HCI pH 8.0, 1 mM EDTA pH 8.0. q) The DNA concentration and purity were calculated by measuring the absorbance of the solution at 230, 260, 280nm. 15 For DNA extraction from Gram Positive bacteria the protocol was as follows:. a) Using a sterile toothpick bacterial colonies were scraped off the culture plate into a sterile 1.5 ml centrifuge tube. b) 180 pl of 20 mg/ml Lysozyme (Sigma) and 200 pg of Lysostaphin (Sigma) was added 20 to each sample and the samples were mixed gently to resuspend the bacterial colonies. c) The samples were incubated at 37 0 C for 30 minutes to degrade the cell wall. d) The samples were then processed and the DNA extracted according to the QIAamp DNA mini kit protocol for Gram positive bacteria. 25 DNA extraction from Cytology samples from patients. a) The sample was shaken vigorously by hand to resuspend any sedimented cells and to ensure the homogeneity of the solution. b) 4 ml of the resuspended cells were transferred to a 15 ml Costar centrifuge tube. 30 c) The tubes were centrifuged in a swing-out bucket rotor at 3000 x g for 15 minutes. d) The supernatant was carefully decanted and discarded without disturbing the pelleted cellular material. e) The pelleted cells were resuspended in 200 pl of lysis buffer (100 mM Tris/HCI pH 8.0, 2 mM EDTA pH 8.0, 0.5% SDS, 0.5% Triton-X-100) and mixed well until the 35 solution was homogeneous.
WO 2007/140506 PCT/AU2006/000755 33 f) 80 pl of the sample was transferred to a 96 well sample preparation plate g) 20 pl of Proteinase K was added and the solution incubated at 550C for 1 hour (this procedure results in cell lysis) DNA extraction from urine samples 5 DNA was extracted from a starting volume of 1 ml of urine according to the QlAamp UltraSensTM Virus Handbook. Bisulfite treatment of DNA samples Bisulfite treatment was carried out according the MethylEasy T M High Throughput 10 DNA bisulfite modification kit (Human Genetic Signatures, Australia) see also below.. Surprisingly, it has been found by the present inventors that there is no need to separate the microbial DNA from other sources of nucleic acids, for example when there is microbial DNA in a sample of human cells. The treatment step can be used for an vast mixture of different DNA types and yet a microbial-specific nucleic acid can be still 15 identified by the present invention. It is estimated that the limits of detection in a complex DNA mixtures are that of the limits of standard PCR detection which can be down to a single copy of a target nucleic acid molecule. Samples 20 Any suitable sample can be used for the present invention. Examples include, but not limited to, microbial cultures, clinical samples, veterinary samples, biological fluids, tissue culture samples, environmental samples, water samples, effluent. As the present invention is adaptable for detecting any microorganism, this list should not be considered as exhaustive. 25 Kits The present invention can be implemented in the form of various kits, or combination of kits and instantiated in terms of manual, semi automated or fully robotic platforms. In a preferred form, the MethyEasy T M or HighThroughput MethylEasy T M kits 30 (Human Genetic Signatures Pty Ltd, Australia) allow conversion of nucleic acids in 96 or 384 plates using a robotic platform such as EpMotion.
WO 2007/140506 PCT/AU2006/000755 34 Bisulfite treatment An exemplary protocol for effective bisulfite treatment of nucleic acid is set out below. The protocol results in retaining substantially all DNA treated. This method is also referred to herein as the Human Genetic Signatures (HGS) method. It will be 5 appreciated that the volumes or amounts of sample or reagents can be varied. Preferred method for bisulfite treatment can be found in US 10/428310 or PCT/AU2004/000549 incorporated herein by reference. To 2 pg of DNA, which can be pre-digested with suitable restriction enzymes if so desired, 2 pl (1/10 volume) of 3 M NaOH (6g in 50 ml water, freshly made) was added in 10 a final volume of 20 pl. This step denatures the double stranded DNA molecules into.a single stranded form, since the bisulfite reagent preferably reacts with single stranded molecules. The mixture was incubated at 370C for 15 minutes. Incubation at temperatures above room temperature can be used to improve the efficiency of denaturation. 15 After the incubation, 208 pl 2 M Sodium Metabisulfite (7.6 g in 20 ml water with 416 ml 10 N NaOH; BDH AnalaR #10356.4D; freshly made) and 12 pl of 10 mM Quinol (0.055 g in 50 ml water, BDH AnalR #103122E; freshly made) were added in succession. Quinol is a reducing agent and helps to reduce oxidation of the reagents. Other reducing agents can also be used, for example, dithiothreitol (DTT), mercaptoethanol, quinone 20 (hydroquinone), or other suitable reducing agents. The sample was overlaid with 200 pl of mineral- oil. The overlaying of mineral oil prevents evaporation and oxidation of the reagents but is not essential. The sample was then incubated overnight at 550C. Alternatively the samples can be cycled in a thermal cycler as follows: incubate for about 4 hours or overnight as follows: Step 1, 550C / 2 hr cycled in PCR machine; Step 2, 950C 25 / 2 min. Step 1 can be performed at any temperature from about 370C to about 900C and can vary in length from 5 minutes to 8 hours. Step 2 can be performed at any temperature from about 700C to about 990C and can vary in length from about 1 second to 60 minutes, or longer. After the treatment with Sodium Metabisulfite, the oil was removed, and 1 pl tRNA 30 (20 mg/ml) or 2 pl glycogen were added if the DNA concentration was low. These additives are optional and can be used to improve the yield of DNA obtained by co precipitating with the target DNA especially when the DNA is present at low concentrations. The use of additives as carrier for more efficient precipitation of nucleic acids is generally desired when the amount nucleic acid is <0.5 pg.
WO 2007/140506 PCT/AU2006/000755 35 An isopropanol cleanup treatment was performed as follows: 800 pl of water were added to the sample, mixed and then 1 ml isopropanol was added. The water or buffer reduces the concentration of the bisulfite salt in the reaction vessel to a level at which the salt will not precipitate along with the target nucleic acid of interest. The 5 dilution is generally about 1/4 to 1/1000 so long as the salt concentration is diluted below a desired range, as disclosed herein. The sample was mixed again and left at 40C for a minimum of 5 minutes. The sample was spun in a microfuge for 10-15 minutes and the pellet was washed 2x with 70% ETOH, vortexing each time. This washing treatment removes any residual salts 10 that precipitated with the nucleic acids. The pellet Was allowed to dry and then resuspended in a suitable volume of TIE (10 mM Tris/0.1 mM EDTA) pH 7.0-12.5 such as 50 pl. Buffer at pH 10.5 has been found to be particularly effective. The sample was incubated at 370C to 950C for 1 min to 96 hr, as needed to suspend the nucleic acids. 15 Another example of bisulfite treatment can be found in WO 2005021778 (incorporated herein by reference) which provides methods and materials for conversion of cytosine to uracil. In some embodiments, a nucleic acid, such as gDNA, is reacted with bisulfite and a polyamine catalyst, such as a triamine or tetra-amine: Optionally, the bisulfite comprises magnesium bisulfite. In other embodiments, a nucleic acid is reacted 20 with magnesium bisulfite, optionally in the presence of a polyamine catalyst and/or a quaternary amine catalyst. Also provided are kits that can be used to carry out methods of the invention. It will be appreciated that these methods would also be suitable for the present invention in the treating step. 25 Amplification PCR amplifications were performed in 25 pl reaction mixtures containing 2 pl of bisulfite-treated genomic DNA, using the Promega PCR master mix, 6 ng/pl of each of the primers. Strand-specific nested primers are used for amplification. 1 st round PCR amplifications were carried out using PCR primers 1 and 4 (see below). Following 1 st 30 round amplification, 1 pl of the amplified material was transferred to 2 nd round PCR premixes containing PCR primers 2 and 3 and amplified as previously described. Samples of PCR products were amplified in a ThermoHybaid PX2 thermal cycler under the conditions: 1 cycle of 950C for 4 minutes, followed by 30 cycles of 950C for 1 minute, 500C for 2 minutes and 720C for 2 minutes; 1 cycle of 720C for 10 minutes.
WO 2007/140506 PCT/AU2006/000755 36 #1 0-14 #4 #2 #3 5 Multiplex amplification If multiplex amplification is required for detection, the following methodology can be carried out. One pl of bisulfite treated DNA is added to the following components in a 25 pl reaction volume, xl Qiagen multiplex master mix, 5-100 ng of each 1 st round INA or 10 oligonucleotide primer 1.5- 4.0 mM MgSO 4 , 400 uM of each dNTP and 0.5-2 unit of the polymerase mixture. The components are then cycled in a hot lid thermal cycler as follows. Typically there can be up to 200 individual primer sequences in each amplification reaction Step 1 940C 15 minute 1 cycle 15 Step 2 94 0 C 1 minute 500C 3 minutes 35 cycles 680C 3 minutes Step 3 680C 10 minutes 1 cycle A second round amplification is then performed on a 1 pl aliquot of the first round 20 amplification that is transferred to a second round reaction tube containing the enzyme reaction mix and appropriate second round primers. Cycling is then performed as above. Primers Any suitable PCR primers can be used for the present invention. A primer 25 typically has a complementary sequence to a sequence which will be amplified. Primers are typically oligonucleotides but can be oligonucleotide analogues. Probes The probe may be any suitable nucleic acid molecule or nucleic acid analogue. 30 Examples include, but not limited to, DNA, RNA, locked nucleic acid (LNA), peptide nucleic acid (PNA), MNA, altritol nucleic acid (ANA), hexitol nucleic acid (HNA), WO 2007/140506 PCT/AU2006/000755 37 intercalating nucleic acid (INA), cyclohexanyl nucleic acid (CNA) and mixtures thereof and hybrids thereof, as well as phosphorous atom modifications thereof, such as but not limited to phosphorothioates, methyl phospholates, phosphoramidites, phosphorodithiates, phosphoroselenoates, phosphotriesters and phosphoboranoates. 5 Non-naturally occurring nucleotides include, but not limited to the nucleotides comprised within DNA, RNA, PNA, INA, HNA, MNA, ANA, LNA, CNA, CeNA, TNA, (2'-NH)-TNA, (3'-NH)-TNA, a-L-Ribo-LNA, a-L-Xylo-LNA, P-D-Xylo-LNA, ca-D-Ribo-LNA, [3.2.1]-LNA, Bicyclo-DNA, 6-Amino-Bicyclo-DNA, 5-epi-Bicyclo-DNA, a-Bicyclo-DNA, Tricyclo-DNA, Bicyclo[4.3.0]-DNA, Bicyclo[3.2.1]-DNA, Bicyclo[4.3.0]amide-DNA, J3-D-Ribopyranosyl 10 NA, -L-Lyxopyranosyl-NA, 2'-R-RNA, a-L-RNA or a-D-RNA, P3-D-RNA. In addition non phosphorous containing compounds may be used for linking to nucleotides such as but not limited to methyliminomethyl, formacetate, thioformacetate and linking groups comprising amides. In particular nucleic acids and nucleic acid analogues may comprise one or more intercalator pseudonucleotides. 15 Preferably, the probes are DNA or DNA oligonucleotides containing one or more internal IPNs forming INA. Electrophoresis Electrophoresis of samples was performed according to the E-gel system user 20 guide (www.invitrogen.doc). Detection methods Numerous possible detection systems exist to determine the status of the desired sample. It will be appreciated that any known system or method for detecting nucleic 25 acid molecules could be used for the present invention. Detection systems include, but not limited to: I. Hybridization of appropriately labelled DNA to a micro-array type device which could select for 10->200,000 individual components. The arrays could be composed of either INAs, PNAs or nucleotide or modified nucleotides arrays onto 30 any suitable solid surface such as glass, plastic, mica, nylon, bead, magnetic bead, fluorescent bead or membrane; II. Southern blot type detection systems; Ill. Standard PCR detection systems such as agarose gel, fluorescent read outs such as Genescan analysis. Sandwich hybridisation assays, DNA staining WO 2007/140506 PCT/AU2006/000755 38 reagents such as ethidium bromide, Syber green, antibody detection, ELISA plate reader type devices, fluorimeter devices; IV. Real-Time PCR quantitation of specific or multiple genomic amplified fragments or any variation on that. 5 V. Any of the detection systems outlined in the WO 2004/065625 such as fluorescent beads, enzyme conjugates, radioactive beads and the like; VI. Any other detection system utilizing an amplification step such as ligase chain reaction or Isothermal DNA amplification technologies such as Strand Displacement Amplification (SDA). 10 VII. Multi-photon detection systems. VIII. Electrophoresis and visualisation in gels. IX. Any detection platform used or could be used to detect nucleic acid. Intercalating nucleic acids 15 Intercalating nucleic acids (INA) are non-naturally occurring polynucleotides which can hybridize to nucleic acids (DNA and RNA) with sequence specificity. INA are candidates as alternatives/substitutes to nucleic acid probes in probe-based hybridization assays because they exhibit several desirable properties. INA are polymerswhich hybridize to nucleic acids to form hybrids which are more thermodynamically stable than 20 a corresponding naturally occurring nucleic acid/nucleic acid complex. They are not substrates for the enzymes which are known to degrade peptides or nucleic acids. Therefore, INA should be more stable in biological samples, as well as, have a longer shelf-life than naturally occurring nucleic acid fragments. Unlike nucleic acid hybridization which is very dependent on ionic strength, the hybridization of an INA with a 25 nucleic acid is fairly independent of ionic strength and is favoured at low ionic strength under conditions which strongly disfavour the hybridization of naturally occurring nucleic acid to nucleic acid. The binding strength of INA is dependent on the number of intercalating groups engineered into the molecule as well as the usual interactions from hydrogen bonding between bases stacked in a specific fashion in a double stranded 30 structure. Sequence discrimination is more efficient for INA recognizing DNA than for DNA recognizing DNA. Preferably, the INA is the phosphoramidite of (S)-1 -O-(4,4' dimethoxytriphenylmethyl)-3-O-(1 -pyrenylmethyl)-glycerol. INA are synthesized by adaptation of standard oligonucleotide synthesis 35 procedures in a format which is commercially available. Full definition of INA and their WO 2007/140506 PCT/AU2006/000755 39 synthesis can be found in WO 03/051901, WO 03/052132, WO 03/052133 and WO 03/052134 (Unest A/S, assigned to Human Genetic Signatures Pty Ltd, Australia) incorporated herein by reference. There are indeed many differences between INA probes and standard nucleic 5 acid probes. These differences can be conveniently broken down into biological, structural, and physico-chemical differences. As discussed above and below, these biological, structural, and physico-chemical differences may lead to unpredictable results when attempting to use INA probes in applications were nucleic acids have typically been employed. This non-equivalency of differing compositions is often observed in the 10 chemical arts. With regard to biological differences, nucleic acids are biological materials that play a central role in the life of living species as agents of genetic transmission and expression. Their in vivo properties are fairly well understood. INA, however, is a recently developed totally artificial molecule, conceived in the minds of chemists and 15 made using synthetic organic chemistry. It has no known biological function. Structurally, INA also differs dramatically from nucleic acids. Although both can employ common nucleobases (A, C, G, T, and U), the composition of these molecules is structurally diverse. The backbones of RNA, DNA and INA are composed of repeating phosphodiester ribose and 2-deoxyribose units. INA differ from DNA or RNA in having 20 one or more large flat molecules attached via a linker molecule(s) to the polymer. The flat molecules intercalate between bases in the complementary DNA stand opposite the INA in a double stranded structure. The physico/chemrical differences between INA and DNA or RNA are also substantial. INA binds to complementary DNA more rapidly than nucleic acid probes 25 bind to the same target sequence. Unlike DNA or RNA fragments, INA bind poorly to RNA unless the intercalating groups are located in terminal positions. Because of the strong interactions between the intercalating groups and bases on the complementary DNA strand, the stability of the INA/DNA complex is higher than that of an analogous DNA/DNA or RNA/DNA complex. 30 Unlike other nucleic acids such as DNA or RNA fragments or PNA, INA do not exhibit self aggregation or binding properties. As INA hybridize to nucleic acids with sequence specificity, INA are useful candidates for developing probe-based assays and are particularly adapted for kits and screening assays. INA probes, however, are not the equivalent of nucleic acid probes.
WO 2007/140506 PCT/AU2006/000755 40 Consequently, any method, kits or compositions which could improve the specificity, sensitivity and reliability of probe-based assays would be useful in the detection, analysis and quantitation of DNA containing samples. INA have the necessary properties for this purpose. 5 RESULTS To demonstrate the present invention, derivative and simplified nucleic acid for Hepatitis C la are shown in Figures 2 to 6. Top and bottom strands of Hepatitis C la native genome are shown in Figures 1 and 2, respectively. Figure 3 and 4 show 10 derivative nucleic acid where all cytosines have been replaced by uracils in the top and bottom strands, respectively. Figure 5 and 6 show modified nucleic acid where all uracils in the derivative nucleic acid have been replaced by thymines to form modified Hepatitis C l a nucleic acid of top and bottom strands, respectively. As can be seen from Figures 2 to 6, essentially four new artificial genomes have 15 been created for Hepatitis C l a which can be used for detection or to obtain suitable targets. The derivative and modified nucleic acid do not exist in nature but are typically generated by bisulphite treatment (derivative) and amplification (modified). The invention therefore is directed at novel nucleic acid molecules generated from naturally occurring microbial nucleic acid which has novel and desirable uses. 20 Table 1 shows the list of derivative and modified microbial nucleic acid sequences according to the invention that are provided in accompanying Sequence Listings (numbered in numerical order). As the size and total number of sequences are extremely large, paper copies have not been provided in the present specification. All sequences are however, incorporated herein by reference. 25 Table 1 Sequence Microorganism Sequence SEQ ID SListing# NO: 1 Acinetobacter Acinetobacter U* 1 Acinetobacter T* 2 Acinetobacter U RC* 3 Acinetobacter T RC* 4 2 Bacillus Bacillus Anthracis U 1 Bacillus Anthracis T 2 Bacillus Anthracis U RC 3 WO 2007/140506 PCT/AU2006/000755 41 Sequence Microorganism Sequence SEQ ID Listing # NO: Bacillus Anthracis T RC 4 Bacilus Cereus U 5 Baclus Cereus T 6 Baclus Cereus U RC 7 Bacllus Cereus T RC 8 Bacillus Subtilis U 9 Bacillus Subtilis T 10 Bacillus Subtilis U RC 11 Bacillus Subtilis T RC 12 3 Bacteriodes Bacteriodes fragilis U 1 Bacteriodes fragilis T 2 Bacteriodes fragilis U RC 3 Bacteriodes fragilis T RC 4 4 Bartonella Bartonella henselae U 1 Bartonella henselae T 2 Bartonella henselae U RC 3 Bartonella henselae T RC 4 Bartonella quintana U 5 Bartonella quintana T 6 Bartonella quintana U RC 7 Bartonella quintana T RC 8 5 Bordetella Bordetella pertussis U 1 Bordetella pertussis T 2 Bordetella pertussis U RC 3 Bordetella pertussis T RC 4 6 Borrelia Borrelia burgdorferi U 1 Borrelia burgdorferi T 2 Borrelia burgdorferi U RC 3 Borrelia burgdorferi T RC 4 7 Brucella Brucella melitensis Chrl U 1 Brucella melitensis Chrl T 2 Brucella melitensis Chrl U RC 3 Brucella melitensis Chrl T RC 4 Brucella melitensis Chr2 U 5 Brucella melitensis Chr2 T 6 Brucella melitensis Chr2 U RC 7 Brucella melitensis Chr2 T RC 8 8 Campylobacter Campylobacter jejuni U 1 WO 2007/140506 PCT/AU2006/000755 42 Sequence Microorganism Sequence SEQ ID Listing # NO: Campylobacter jejuni T 2 Campylobacterjejuni U RC 3 Campylobacter jejuni T RC 4 9 Chlamydia Chlamydia Pneumoniae U 1 Chlamydia Pneumoniae T 2 Chlamydia Pneumoniae U RC 3 Chlamydia Pneumoniae T RC 4 Chlamydia trachomatis U 5 Chlamydia trachomatis T 6 Chlamydia trachomatis U RC 7 _ _Chlamydia trachomatis T RC 8 10 Clostridium Clostridium botulinum Toxin A U 1 Clostridium botulinum Toxin A T 2 Clostridium botulinum Toxin A U RC 3 Clostridium botulinum Toxin A T RC 4 Clostridium botulinum Toxin B U 5 Clostridium botulinum Toxin B T 6 Clostridium botulinum Toxin B U RC 7 Clostridium botulinum Toxin B T RC 8 Clostridium botulinum Toxin C U 9 Clostridium botulinum Toxin C T 10 Clostridium botblinum Toxin C U RC 11 Clostridium botulinum Toxin C T RC 12 Clostridium botulinum Toxin D U 13 Clostridium botulinum Toxin D T 14 Clostridium botulinum Toxin D U RC 15 Clostridium botulinum Toxin D T RC 16 Clostridium botulinum Toxin E U 17 Clostridium botulinum Toxin E T 18 Clostridium botulinum Toxin E F RC 19 Clostridium botulinum Toxin E T RC 20 Clostridium botulinum Toxin F U 21 Clostridium botulinum Toxin F T 22 Clostridium botulinum Toxin F U RC 23 Clostridium botulinum Toxin F T RC 24 Clostridium dififcile U 25 Clostridium dififcile T 26 Clostridium dififcile U RC 27 Clostridium dififcile T RC 28 WO 2007/140506 PCT/AU2006/000755 43 Sequence Microorganism Sequence SEQ ID Listing # NO: Clostridium perfringens U 29 Clostridium perfringens T 30 Clostridium perfringens U RC 31 Clostridium perfringens T RC 32 Clostridium tetani U 33 Clostridium tetani T 34 Clostridium tetani U RC 35 Clostridium tetani T RC 36 Clostridium botulinum U 37 Clostridium botulinum T 38 Clostridium botulinum U RC 39 Clostridium botulinum T RC 40 11 Corynebacterium Corynebacterium diptheriae U I1 Corynebacterium diptheriae T 2 Corynebacterium diptheriae U RC 3 Corynebacterium diptheriae T RC 4 Corynebacterium jeikeium U 5 Corynebacterium jeikeium T 6 Corynebacterium jeikeium U RC 7 Corynebacterium jeikelum T RC 8 12 Ecoli Ecoli U 1 Ecoli T 2 Ecoli U RC 3 Ecoli T RC 4 13 Ehrlichia Ehrlichia chaffeensis U 1 Ehrlichia chaffeensis T 2 Ehrlichia chaffeensis U RC 3 Ehrlichia chaffeensis T RC 4 14 Enterococcus Enterococcus faecalis U 1 Enterococcus faecalis T 2 Enterococcus faecalis U RC 3 Enterococcus faecalis T RC 4 15 Fusobacterium Fusobacterium nucleatum U 1 Fusobacterium nucleatum T 2 Fusobacterium nucleatum U RC 3. Fusobacterium nucleatum T RC 4 16 Haemophilus Haemophilus ducreyi U 1 Haemophilus ducreyi T 2 Haemophilus ducreyi U RC 3 WO 2007/140506 PCT/AU2006/000755 44 Sequence Microorganism Sequence SEQ ID Listing # NO: Haemophilus ducreyi T RC 4 Haemophilus influenzae U 5 Haemophilus influenzae T 6 Haemophilus influenzae U RC 7 Haemophilus influenzae T RC 8 17 Helicobacter Helicobacter pylori U 1 Helicobacter pylori T 2 Helicobacter pylori U RC 3 Helicobacter pylori T RC 4 18 Legionella Legionella pneumophila U 1 Legionella pneumophila T 2 Legionella pneumophila U RC 3 Legionella pneumophila T RC 4 19 Leptospira Leptospira interrogans Chr I U 1 Leptospira interrogans Chr I T 2 Leptospira interrogans Chr I U RC 3 Leptospira interrogans Chr I T RC 4 Leptospira interrogans Chr II U 5 Leptospira interrogans Chr II T , 6 Leptospira interrogans Chr II U RC 7 Leptospira interrogans Chr II T RC 8 20 Listeria Listeria monocytogenes U 1 Listeria monocytogenes T 2 Listeria monocytogenes U RC 3 Listeria monocytogenes T RO 4 21 Mycobacterium Mycobacterium avium U 1 Mycobacterium avium T 2 Mycobacterium avium U RC 3 Mycobacterium avium T RC 4 Mycobacterium leprae U 5 Mycobacterium leprae T 6 Mycobacterium leprae U RC 7 Mycobacterium leprae T RC 8 Mycobacterium tuberculosis U 9 Mycobacterium tuberculosis T 10 Mycobacterium tuberculosis U RC 11 Mycobacterium tuberculosis T RC 12 22 Mycoplasma Mycoplasma pneumoniae U 1 Mycoplasma pneumoniae T 2 WO 2007/140506 PCT/AU2006/000755 45 Sequence Microorganism Sequence SEQ ID Listing # NO: Mycoplasma pneumoniae U RC 3 Mycoplasma pneumoniae T RC 4 23 Neisseria Neisseria gonorrhopae U 1 Neisseria gonorrhoeae T 2 Neisseria gonorrhoeae U RC 3 Neisseria gonorrhoeae T RC 4 Neisseria meningitides sero A U 5 Neisseria meningitides sero A T 6 Neisseria meningitides sero A U RC 7 Neisseria meningitides sero A T RC 8 Niesseria meningitidis sero B U 9 Niesseria meningitidis sero B T 10 Niesseria meningitidis sero B U RC 11 Niesseria meningitidis sero B T RC 12 24 Nocardia Nocardia farcinica U 1 Nocardia farcinica T 2 Nocardia farcinica U RC 3 Nocardia farcinica T RC 4 25 Pseudomonas Pseudomonas aeruginosa U 1 Pseudomonas aeruginosa T 2 Pseudomonas aeruginosa U RC 3 Pseudomonas aeruginosa T RC 4 26 Rickettsia Rickettsia prowazekii U 1 Rickettsia prowazekii T 2 Rickettsia prowazekii U RC 3 Rickettsia prowazekii T RC 4 Rickettsia typhi U 5 Rickettsia typhi T 6 Rickettsia typhi U RC 7 Rickettsia typhi T RC 8 Rickettsia conorii U 9 Rickettsia conorii T 10 Rickettsia conorii U RC 11 Rickettsia conorii T RC 12 27 Salmonella Salmonella enterica U 1 Salmonella enterica T 2 Salmonella enterica U RC 3 Salmonella enterica T RC 4 Salmonella typhimurium U 5 WO 2007/140506 PCT/AU2006/000755 46 Sequence Microorganism Sequence SEQ ID Listing # NO: Salmonella typhimurium T 6 Salmonella typhimurium U RC 7 Salmonella typhimurium T RC 8 28 Serratia Serratia marscesens U 1 Serratia marscesens T 2 Serratia marscesens U RC 3 Serratia marscesens T RC 4 29 Shigella Shigella flexneri U 1 Shigella flexneri T 2 Shigella flexneri U RC 3 Shigella flexneri T RC 4 Shigella boydii U 5 Shigella boydli T 6 Shigella boydii U RC 7 Shigella boydii T RC 8 Shigella dysenteriae U 9 Shigella dysenteriae T 10 Shigella dysenteriae U RC 11 Shigella dysenteriae T RC 12 30 Staphylococcus Staphylococcus aureus U 1 Staphylococcus aureus T 2 Staphylococcus aureus U RC 3 Staphylococcus aureus T RC 4 Staphylococcus epidermidis U 5 Staphylococcus epidermidis T 6 Staphylococcus epidermidis U RC 7 Staphylococcus epidermidis T RC 8 Staphylococcus haemolyticus U 9 Staphylococcus haemolyticus T 10 Staphylococcus haemolyticus U RC 11 Staphylococcus haemolyticus T RC 12 Staphylococcus enterotoxin A U 13 Staphylococcus enterotoxin AT 14 Staphylococcus enterotoxin A U RC 15 Staphylococcus enterotoxin AT RC 16 Staphylococcus enterotoxin B U 17 Staphylococcus enterotoxin B T 18 Staphylococcus enterotoxin A B RC 19 Staphylococcus enterotoxin A B RC 20 WO 2007/140506 PCT/AU2006/000755 47 Sequence Microorganism Sequence SEQ ID Listing # NO: Staphylococcus enterotoxin CU 21 Staphylococcus enterotoxin C T 22 Staphylococcus enterotoxin C U RC 23 Staphylococcus enterotoxin C T RC 24 Staphylococcus enterotoxin Cl U 25 Staphylococcus enterotoxin Cl T 26 Staphylococcus enterotoxin Cl U RC 27 Staphylococcus enterotoxin C1 T RC 28 Staphylococcus enterotoxin C3 U 29 Staphylococcus enterotoxin C3 T 30 Staphylococcus enterotoxin C3 U RC 31 Staphylococcus enterotoxin C3 T RC 32 Staphylococcus enterotoxin D U 33 Staphylococcus enterotoxin D T 34 Staphylococcus enterotoxin D U RC 35 Staphylococcus enterotoxin D T RC 36 Staphylococcus enterotoxin E U 37 Staphylococcus enterotoxin E T 38 Staphylococcus enterotoxin E U RC 39 Staphylococcus enterotoxin E T RC 40 Staphylococcus enterotoxin G U 41 Staphylococcus enterotoxin G T 42 Staphylococcus enterotoxin G U RC 43 Staphylococcus enterotoxin G T RC 44 Staphylococcus enterotoxin H U 45 Staphylococcus enterotoxin H T 46 Staphylococcus enterotoxin H U RC 47 Staphylococcus enterotoxin H T RC 48 Staphylococcus enterotoxin J&R U 49 Staphylococcus enterotoxin J&R T 50 Staphylococcus enterotoxin J&R U RC 51 Staphylococcus enterotoxin J&R T RC 52 31 Streptococcus Streptococcus agalactiae U 1 Streptococcus agalactiae T 2 Streptococcus agalactiae U RC 3 Streptococcus agalactiae T RC 4 Streptococcus mutans U 5 Streptococcus mutans T 6 Streptococcus mutans U RC 7 WO 2007/140506 PCT/AU2006/000755 48 Sequence Microorganism Sequence SEQ ID Listing # NO: Streptococcus mutans T RC 8 Streptococcus pneumoniae U 9 Streptococcus pneumoniae T 10 Streptococcus pneumoniae U RC 11 Streptococcus pneumoniae T RC 12 Streptococcus pyrogene U 13 Streptococcus pyrogene T 14 Streptococcus pyrogene U RC 15 Streptococcus pyrogene T RC 16 32 Streptomyces Streptomyces coelicolor U 1 Streptomyces coelicolor T 2 Streptomyces coelicolor U RC 3 Streptomyces coelicolor T RC 4 33 Treponema Treponema pallidumU 1 Treponema pallidumT 2 Treponema pallidumU RC 3 Treponema pallidumT RC 4 34 Tropheryma Tropheryma whippelii U 1 Tropheryma whippelii T 2 Tropheryma whippelii U RC 3 Tropheryma whippelii T RC 4 35 Plasmodium Plasmodium falciparum mitochondrion U 1 Plasmodium falciparum mitochondrion T 2 Plasmodium falciparum mitochondrion U RC 3 Plasmodium falciparum mitochondrion T RC 4 Plasmodium falciparum Chrl U 5 Plasmodium falciparum Chri T 6 Plasmodium falciparum Chrl U RC 7 Plasmodium falciparum Chrl T RC 8 Plasmodium falciparum Chr2 U 9 Plasmodium falciparum Chr2 T 10 Plasmodium falciparum Chr2 U RC 11 Plasmodium falciparum Chr2 T RC 12 Plasmodium falciparum Chr3 U 13 Plasmodium falciparum Chr3 T 14 Plasmodium falciparum Chr3 U RC 15 Plasmodium falciparum Chr3 T RC 16 Plasmodium falciparum Chr4 U 17 Plasmodium falciparum Chr4 T 18 WO 2007/140506 PCT/AU2006/000755 49 Sequence Microorganism Sequence SEQ ID Listing # NO: Plasmodiu.m falciparum Chr4 U RC 19 Plasmodium falciparum Chr4 T RC 20 Plasmodium falciparum Chr5 U 21 Plasmodium falciparum Chr5 T 22 Plasmodium falciparum Chr5 U RC 23 Plasmodium falciparum Chr5 T RC 24 Plasmodium falciparum Chr6 U 25 Plasmodium falciparum Chr6 T 26 Plasmodium falciparum Chr6 U RC 27 Plasmodium falciparum Chr6 T RC 28 Plasmodium falciparum Chr7 U 29 Plasmodium falciparum Chr7 T 30 Plasmodium falciparum Chr7 U RC 31 Plasmodium falciparum Chr7 T RC 32 Plasmodium falciparum Chr8 U 33 Plasmodium falciparum Chr8 T 34 Plasmodium falciparum Chr8 U RC 35 Plasmodium falciparum Chr8 T RC 36 Plasmodium falciparum Chr9 U 37 Plasmodium falciparum Chr9 T 38 Plasmodium falciparum Chr9 U RC 39 Plasmodium falciparum Chr9 T RC 40 Plasmodium falciparum ChrlO U 41 Plasmodium falciparum Chrl0 T 42 Plasmodium falciparum Chrl0 U RC 43 Plasmodium falciparum ChrlO T RC 44 Plasmodium falciparum Chr 11 U 45 Plasmodium falciparum Chr 11 T 46 Plasmodium falciparum Chr 11 U RC 47 Plasmodium falciparum Chrl 1 T RC 48 Plasmodium falciparum Chrl2 U 49 Plasmodium falciparum ChrI 2 T 50 Plasmodium falciparum Chrl2 U RC 51 Plasmodium falciparum Chrl 2 T RC 52 Plasmodium falciparum Chrl3 U 53 Plasmodium falciparum Chrl 3 T 54 Plasmodium falciparum Chrl 3 U RC 55 Plasmodium falciparum Chrl 3 T RC 56 Plasmodiumr falciparum Chrl4 U 57 WO 2007/140506 PCT/AU2006/000755 50 Sequence Microorganism Sequence SEQ ID Listing # NO: Plasmodium falciparum Chrl4 T 58 Plasmodium falciparum Chr14 U RC 59 Plasmodium falciparum Chrl4 T RC 60 36 Aspergillus Aspergillus niger mitochondrion U 1 Aspergillus niger mitochondrion T 2 Aspergillus niger mitochondrion U RC 3 Aspergillus niger mitochondrion T RC 4 Aspergillus tubingensis mito U 5 Aspergillus tubingensis mito T 6 Aspergillus tubingensis mito U RC 7 Aspergillus tubingensis mito T RC 8 37 Candida albicans mito U 1 Candida albicans mito T 2 Candida albicans mito U RC 3 Candida albicans mito T RC 4 Candida glabrate mito U 5 Candida glabrate mito T 6 Candida glabrate mito U RC 7 Candida glabrate mito T RC 8 Candida metapsilosis mito U 9 Candida metapsilosis mito T 10 Candida metapsilosis mito U RC 11 Candida metapsilosis mito T RC 12 Candida orthopsilosis mito U 13 Candida orthopsilosis mito T 14 Candida orthopsilosis mito U RC 15 Candida orthopsilosis mito T RC 16 Candida parapsilosis mito U 17 Candida parapsilosis mito T 18 Candida parapsilosis mito U RC 19 Candida parapsilosis mito T RC 20 Candida stellata mito U 21 Candida stellata mito T 22 Candida stellata mito U RC 23 Candida stellata mito T RC 24 38 Cryptococcus Cryptococcus neoformans var U 1 Cryptococcus neoformans var T 2 Cryptococcus neoformans var U RC 3 Qryptococcus neoformans var T RC 4 WO 2007/140506 PCT/AU2006/000755 51 Sequence Microorganism Sequence SEQ ID Listing # NO: 39 Paracoccidioides Paracoccidioides brasiliensis mito U 1 Paracoccidioides brasiliensis mito T 2 Paracoccidioides brasiliensis mito U RC 3 Paracoccidioides brasiliensis mito T RC 4 40 Rhizopus Rhizopus oryzae mitochondrion U 1 Rhizopus oryzae mitochondrion T 2 Rhizopus oryzae mitochondrion U RC 3 Rhizopus oryzae mitochondrion T RC 4 41 Francisella Francisella tularensis U 1 Francisella tularensis T 2 Francisella tularensis U RC 3 Francisella tularensis T RC 4 42 VYibrio Vibrio cholerae chromosoame 11 U 1 Vibrio cholerae chromosoame 1.1 T 2 Vibrio cholerae chromosoame II U RC 3 Vibrio cholerae chromosoame II T RC 4 43 Yersinia Yersinia pestis U 1 Yersinia pestis T 2 Yersinia pestis U RC 3 Yersinia pestis T RC 4 44 JC polyomavirus JC polyomavirus U 1 JC polyomavirus T 2 JC polyomavirus U RC 3 JC polyomavirus T RC 4 45 1 2 3 4 46 Andes virus Andes virus segment L U 1 Andes virus segment L T 2 Andes virus segment L U RC 3 Andes virus segment L T RC 4 Andes virus segment M U 5 Andes virus segment M T 6 Andes virus segment M U RC 7 Andes virus segment M T RC 8 Andes virus segment S U 9 Andes virus segment ST 10 Andes virus segment S U RC 11 WO 2007/140506 PCT/AU2006/000755 52 Sequence Microorganism Sequence SEQ ID Listing # NO: Andes virus segment S T RC 12 51 Hepatitis C virus Hepatitis C 1 a T 1 Hepatitis C 2b T 2 Hepatitis C 1b/2k T 3 Hepatitis C 1c T 4 Hepatitis C 2a T 5 Hepatitis C 2b T 6 Hepatitis C 2C T 7 Hepatitis C 2K T 8 Hepatitis C 3a T 9 Hepatitis C 3B T 10 Hepatitis C 3k T 11 Hepatitis C 4a T 12 Hepatitis C Sa T 13 Hepatitis C 6a T 14 Hepatitis C 6b T 15 Hepatitis C 6g T 16 Hepatitis C 6d T 17 Hepatitis C 6h T 18 Hepatitis C 6k T 19 Hepatitis C la T RC 20 Hepatitis C 2b T RC 21 Hepatitis C 1 b/2k T RC 22 Hepatitis C 1c T RC 23 Hepatitis C 2a T RC 24 Hepatitis C 2b T RC 25 Hepatitis C 2C T RC 26 Hepatitis C 2K T RC 27 Hepatitis C 3a T RC 28 WO 2007/140506 PCT/AU2006/000755 53 Sequence Microorganism Sequence SEQ ID Listing # NO: Hepatitis C 3B T RC 29 Hepatitis C 3k T RC 30 Hepatitis C 4a T RC 31 Hepatitis C Sa T RC 32 Hepatitis C 6a T RC 33 Hepatitis C 6b T RC 34 Hepatitis C 6g T RC 35 Hepatitis C 6d T RC 39 Hepatitis C 6h T RC 37 Hepatitis C 6k T RC 38 Hepatitis C la U 39 Hepatitis C 2b U 40 Hepatitis C lb/2k U 41 Hepatitis C 1c U 42 Hepatitis C 2a U 43 Hepatitis C 2b U 44 Hepatitis C 2C U 45 Hepatitis C 2K U 46 Hepatitis C 3a U 47 Hepatitis C 3B U 48 Hepatitis C 3k U 49 Hepatitis C 4a U 50 Hepatitis C 5a U 51 Hepatitis C 6a U 52 Hepatitis C 6b U 53 Hepatitis C 6g U 54 Hepatitis C 6d U 55 Hepatitis C 6h U 56 Hepatitis C 6k U 57 Hepatitis C la U RC 58 WO 2007/140506 PCT/AU2006/000755 54 Sequence Microorganism Sequence SEQ ID Listing # NO: Hepatitis C 2b U RC 59 Hepatitis C I b/2k U RC 60 Hepatitis C Ic U RC 61 Hepatitis C 2a U RC 62 Hepatitis C 2b U RC 63 Hepatitis C 2C U RC 64 Hepatitis C 2K U RC 65 Hepatitis C 3a U RC 66 Hepatitis C 3B U RC 67 Hepatitis C 3k U RC 68 Hepatitis C 4a U RC 69 Hepatitis C 5a U RC 70 Hepatitis C 6a U RC 71 Hepatitis C 6b U RC 72 Hepatitis C 6g U RC 73 Hepatitis C 6d U RC 74 Hepatitis C 6h U RC 75 Hepatitis C 6k U RC 76 Hepatitis C l a genome T 77 Hepatitis C la genome UC 78 52 Hepatitis Hepatitis A 1 Hepatitis B 2 Hepatitis D 3 Hepatitis E 4 Hepatitis GA 5 Hepatitis GB , 6 Hepatitis A RC 7 Hepatitis B RC 8 Hepatitis D RC 9 Hepatitis E RC 10 Hepatitis GA RC 11 Hepatitis GB RC 12 Hepatitis A U 13 Hepatitis B U 14 WO 2007/140506 PCT/AU2006/000755 55 Sequence Microorganism Sequence SEQ ID Listing # NO: Hepatitis D U 15 Hepatitis EU 16 Hepatitis GA U 17 Hepatitis GB U 18 Hepatitis A U RC 19 Hepatitis B U RC 20 Hepatitis D U RC 21 Hepatitis E U RC 22 Hepatitis GA U RC 23 Hepatitis GB U RC 24 53 Human HIV-1 Gp M Clade Al 1 53 Human HIV-1 Gp M Glade A2 2 Immunodeficiency HIV-1 Gp M Clade A2 2 virus (HIV) HIV-1 Gp M Clade B 3 HIV-1 Gp M Clade C. 4 HIV-1 Gp M Clade D 5 HIV-1 Gp M Clade F1 6 HIV-1 Gp M Clade F2 7 HIV-1 Gp M Clade G 8 HIV-1 Gp M Clade H 9 HIV-1 Gp M Clade J 10 HIV-1 Gp M Clade K 11 HIV-1 Gp M CRF A/E 12 HIV-1 Gp M A/G 13 HIV-1 Gp M A/B 14 HIV-1 Gp M A/G/I 15 HIV-1 Gp M F/D 16 HIV-1 Gp M A/G/J 17 HIV-1 Gp M C/B 18 HIV-1 Gp M B/C 19 HIV-1 Gp M A/FI/G 20 HIV-1 Gp M C/D 21 HIV-1 Gp M A/E/J/G 22 HIV-1 Gp M B/F 23 HIV-1 Gp M A, CRF01AE 24 HIV-1 Gp M B/G 25 HIV-1 Gp M CRF01AE/B 26 HIV-1 Gp M A2/D 27 HIV-1 Gp M A,F,G,H,J 28 HIV-1 Gp M A/D/G 29 HIV-1 Gp N 30 HIV-1 Gp O 31 HIV-2 32 HIV-1 Gp M Clade Al RC 33 HIV-1 Gp M Clade A2 RC 34 HIV-1 Gp M Clade B RC 35 HIV-1 Gp M Clade C RC 36 HIV-1 Gp M Clade D RC 37 HIV-1 Gp M Clade F1 RC 38 HIV-1 Gp M Clade F2 RC 39 HIV-1 Gp M Clade G RC 40 HIV-1 Gp M Clade H RC 41 HIV-1 Gp M Clade J RC 42 WO 2007/140506 PCT/AU2006/000755 56 Sequence Microorganism Sequence SEQ ID Listing # NO: HIV-1 Gp M Clade K RC 43 HIV-1 Gp M CRF A/E RC 44 HIV-1 Gp M A/G RC 45 HIV-1 Gp M A/B RC 46 HIV-1 Gp M A/G/I RC 47 HIV-1 Gp M F/D RC 48 HIV-1 Gp M A/G/J RC 49 HIV-1 Gp M C/B RC 50 HIV-1 Gp M B/C RC 51 HIV-1 Gp M A/F/G RC 52 HIV-1 Gp M C/D RC 53 HIV-1 Gp M A/E/J/G RC 54 HIV-1 Gp M B/F RC 55 HIV-1 Gp M A, CRF01AE RC 56 HIV-1 Gp M B/G RC 57 HIV-1 Gp M CRF01AE/B RC 58 HIV-1 Gp M A2/D RC 59 HIV-1 Gp M A,F,G,H,J RC 60 HIV-1 Gp M A/D/G RC 61 HIV-1 Gp N RC. 62 HIV-1 Gp O RC 63 HIV-2 RC 64 HIV-1 Gp M Clade Al U 65 HIV-1 Gp M Clade A2 U 66 HIV-1 Gp M Clade B U 67 HIV-1 Gp M Clade C U 68 HIV-1 Gp M Clade D U 69 HIV-1 Gp M Clade Fl U 70 HIV-1 Gp M Clade F2 U 71 HIV-1 Gp M Clade G U 72 HIV-1 Gp M Clade HU 73 HIV-1 Gp M Clade J U 74 HIV-1 Gp M Clade K U 75 HIV-1 Gp M CRF A/E U 76 HIV-1 Gp M A/G U 77 HIV-1 Gp M A/B U 78 HIV-1 Gp M A/G/I U 79 HIV-1 Gp M F/D U 80 HIV-1 Gp M A/G/J U 81 HIV-1 Gp M C/B U 82 HIV-1 Gp M B/C U 83 HIV-1 Gp M A/F/G U 84 HIV-1 'Gp M C/D U 85 HIV-1 Gp M A/E/J/G U 86 HIV-1 Gp M B/F U 87 HIV-1 Gp M A, CRF01AE U 88 HIV-1 Gp M B/G U 89 HIV-1 Gp M CRF01AE/B U 90 HIV-1 Gp M A2/D U 91 HIV-1 Gp M A,F,G,H,J U 92 HIV-1 Gp M A/D/G U 93 HIV-1 Gp N U 94 HIV-1 Gp OU 95 HIV-2 U 96 HIV-1 Gp M Clade Al U RC 97 HIV-1 Gp M Clade A2 U RC 98 WO 2007/140506 PCT/AU2006/000755 57 Sequence Microorganism Sequence SEQ ID Listing # NO: HIV-1 Gp M Glade B U RC 99 HIV-1 Gp M Glade CU RC 100 HIV-1 Gp M Clade D U RC 101 HIV-1 Gp M Glade Fl U RC 102 HIV-1 Gp M Glade F2 U RC 103 HIV-1 Gp M Glade G U RC 104 HIV-1 Gp M Glade H U RC 105 HIV-1 Gp M Clade J U RC 106 HIV-1 Gp M Glade K U RC 107 HIV-1 Gp M CRF A/E U RC 108 HIV-1 Gp M A/G U RC 109 HIV-1 Gp M A/B U RC 110 HIV-1 Gp M A/G/I U RC 111 HIV-1 Gp M F/D U RC 112 HIV-1 Gp M A/G/J U RC 113 HIV-1 Gp M C/B U RC 114 HIV-1 Gp M B/C U RC 115 HIV-1 Gp M A/F/G U RC 116 HIV-1 Gp M C/D U RC 117 HIV-1 Gp M A/E/J/G U RC 118 HIV-1 Gp M B/F U RC 119 HIV-1 Gp M A,CRF01AE U RC 120 HIV-1 Gp M B/G U RC 121 HIV-1 Gp M CRF01AE/B U RC 122 HIV-1 Gp M A2/D U RC 123 HIV-1 Gp M A,F,G,H,J U RC 124 HIV-1 Gp M A/D/G U RC 125 HIV-1 Gp N U RC 126 HIV-1 Gp O U RC 127 HIV-2 U RC 128 54 Influenza virus Flu A segment 1 1 Flu A segment 2 2 Flu A segment 3 3 Flu A segment 4 4 Flu A segment 5 5 Flu A segment 6 6 Flu A segment 7 7 Flu A segment 8 8 Flu B HA 9 Flu B NP 10 Flu B PB1 T 11 Flu B PB2 T 12 Flu B NB BAT 13 Flu B M1 BM2 T 14 Flu B PAT 15 Flu B NS1 NS2 T 16 Flu B HA HiN1. T 17 Flu B HA H2N2 T 18 Flu B HA H3N2 T 19 Flu B HA H4N2 T 20 Flu B HA H5N1 T 21 Flu B HA H6N2 T 22 Flu B HA H7N1 T 23 Flu B HA H8N4 T 24 Flu B HA H9N2 T 25 WO 2007/140506 PCT/AU2006/000755 58 Sequence Microorganism Sequence SEQ ID Listing # NO: Fly B HA H10ON7 T 26 Flu B HA H11 N2 T 27 Flu B HA H12N1 T 28 Flu B HA H13N2 T 29 Flu B HA H14 T 30 Flu B HA H15N2 T 31 Flu B HA H16N3 T 32 Flu B NA H1N1 T 33 Flu B NA H5N1 T 34 Flu B NA H3N2 T 35 Flu B NA H3N3 T 36 Flu B NA H8N4 T 37 Flu B NA H6N5 T 38 Flu B NA H4N6 T 39 Flu B NA H7N7 T 40 Flu B NA H3N8 T 41 Flu B NA H2N9 T 42 Flu A segment 1 T RC 43 Flu A segment 2 T RC 44 Flu A segment 3 T RC 45 Flu A segment 4 T RC 46 Flu A segment 5 T RC 47 Flu A segment 6 T RC 48 Flu A segment 7 T RC 49 Flu A segment 8 T RC 50 Flu B HAT RC 51 Flu B NP T RC 52 Flu B PB1 T RC 53 Flu B PB2 T RC 54 Flu B NB BA T RC 55 Flu B M1 BM2 T RC 56 Flu B PAT RC 57 Flu B NS1 NS2 T RC 58 Flu B HA H1N1 T RC 59 Flu B HA H2N2 T RC 60 Flu B HA H3N2 T RC 61 Flu B HA H4N2 T RC 62 Flu B HA H5N1 T RC 63 Flu B HA H6N2 T RC 64 Flu B HA H7N1 T RC 65 Flu B HA H8N4 T RC 66 Flu B HA H9N2 T RC 67 Flu B HA H10N7 T RC 68 Flu B HA HIIN2 T RC .
69 Flu B HA H12N1 T RC 70 Flu B HA H13N2 T RC 71 Flu B HA H14 T RC 72 Flu B HA H15N2 T RC 73 Flu B HA H16N3 T RC 74 Flu B NA H1N1 T RC 75 Flu. B NA H5N1 T RC 76 Flu B NA H3N2 T RC 77 Flu B NA H8N4 T RC 78 Flu B NA H6N5 T RC 79 Flu B NA H4N6 T RC 80 Flu B NA H7N7 T RC 81 WO 2007/140506 PCT/AU2006/000755 59 Sequence Microorganism Sequence SEQ ID Listing # NO: Flu B NA H3N8 T RO 82 Flu B NA H2N9 T RC 83 Flu A segment I U 84 Flu A segment 2 U 85 Flu A segment 3 U 86 Flu A segment 4 U 87 Flu A segment 5 U 88 Flu A segment 6 U 89 Flu A segment 7 U 90 Flu A segment 8 U 91 Flu B HA U 92 Flu B NP U 93 Flu B PB1 U 94 Flu B PB2 U 95 Flu B NB NA U 96 Flu B M1 BM2 U 97 Flu B PA U 98 Flu B NS1 NS2 U 99 Flu BHA H1N1 U 100 Flu B HA H2N2 U 101 Flu B HA H3N2 U 102 Flu B HA H4N2 U 103 Flu B HA H5N1 U 104 Flu B HA H6N2 U 105 Flu B HA H7N1 U 106 Flu B HA H8N4 U 107 Flu B HA H9N2 U 108 Flu B HA H10N7 U 109 Flu B HA H11N2 U 110 Flu B HA H12N1 U 111 Flu B HA H13N2 U 112 Flu B HA H14 U 113 Flu B HA H15N2 U 114 Flu B HA H16N3 U 115 Flu B NA H1N1 U 116 Flu B NA H5N1 U 117 Flu B NA H3N2 U 118 Flu B NA H8N4 U 119 Flu B NA H6N5 U 120 Flu B NA H4N6 U 121 Flu B NA H7N7 U 122 Flu B NA H3N8 U 123 Flu B NA H2N9 U 124 Flu A segment 1 U RC 125 Flu A segment 2 U RC 126 Flu A segment 3 U RC 127 Flu A segment 4 U RC 128 Flu A segment 5 U RC 129 Flu A segment 6 U RC 130 Flu A segment 7 U RC 131 Flu A segment 8 U RC 132 Flu B HA U RC 133 Flu B NP U RC 134 Flu B PB1 U RC 135 Flu B PB2 U RC 136 Flu B NB BA U RC 137 WO 2007/140506 PCT/AU2006/000755 60 Sequence Microorganism Sequence SEQ ID Listing # NO: Flu B M1 BM2 U RC, 138 Flu B PA U RC 139 Flu B NS1 N S2 U RC 140 Flu B HA H2N2 U RC 141 Flu B HA H3N2 U RC 142 Flu B HA H4N2 U RC 143 Flu B HA H5N1 U RC 144 Flu B HA H6N2 U RC 145 Flu B HA H7N1 U RC 146 Flu B HA H8N4 U RC 147 Flu B HA H9N2 U RC 148 Flu B HA H10N7 U RC 149 Flu B HA H11N2 U RC 150 Flu B HA H12N1 U RC 151 Flu B HA H13N2 U RC 152 Flu B HA H14 U RC 153 Flu B HA H15N2 U RC 154 Flu B HA H16N3 U RC 155 Flu B NA H1N1 U RC 156 Flu B NA H5N1 U RC 157 Flu B NA H3N2 U RC 158 Flu B NA H6N5 U RC 159 Flu B NA H7N7 U RC 160 Flu B NA H3N8 U RC 161 Flu B NA H2N2 U RC, 162 55 BK BK virus U 1 BK virus T 2 BK virus U RC 3 BK virus T RC 4 56 Barmah Barmah Forest virus U 1 Barmah Forest virus T 2 Barmah Forest virus U RC 3 Barmah Forest virus T RC 4 57 Calicivirus Calicivirus U 1 Calicivirus T 2 Calicivirus U RC 3 Calicivirus T RC 4 58 Colorado tick fever Colorado tick fever segment 1 U 1 virus Colorado tick fever segment 1 T 2 Colorado tick fever segment 1 U RC 3 Colorado tick fever segment 1 T RC 4 Colorado tick fever segment 2 U 5 Colorado tick fever segment 2 T 6 Colorado tick fever segment 2 U RC 7 Colorado tick fever segment 2 T RC 8 Colorado tick fever segment 3 U 9 WO 2007/140506 PCT/AU2006/000755 61 Sequence Microorganism Sequence SEQ ID Listing # NO: Colorado tick fever segment 3 T 10 Colorado tick fever segment 3 U RC 11 Colorado tick fever segment 3 T RC 12 Colorado tick fever segment 4 U 13 Colorado tick fever segment 4 T 14 Colorado tick fever segment 4 U RC 15 Colorado tick fever segment 4 T RC 16 Colorado tick fever segment 5 U 17 Colorado tick fever segment 5 T 18 Colorado tick fever segment 5 U RC 19 Colorado tick fever segment 5 T RC 20 Colorado tick fever segment 6 U 21 Colorado tick fever segment 6 T 22 Colorado tick fever segment 6 U RC 23 Colorado tick fever segment 6 T RC 24 Colorado tick fever segment 7 U 25 Colorado tick fever segment 7 T 26 Colorado tick fever segment 7 U RC 27 Colorado tick fever segment 7 T RC 28 Colorado tick fever segment 8 U 29 Colorado tick fever segment 8 T 30 Colorado tick fever segment 8 U RC 31 Colorado tick fever segment 8 T RC 32 Colorado tick fever segment 9 U 33 Colorado tick fever segment 9 T 34 Colorado tick fever segment 9 U RC 35 Colorado tick fever segment 9 T RC 36 Colorado tidk fever segment 10 U 37 Colorado tick fever segment 10 T 38 Colorado tick fever segment 10 U RC 39 Colorado tick fever segment 10 T RC 40 Colorado tick fever segment 11 U 41 Colorado tick fever segment 11 T 42 Colorado tick fever segment 11 U RC 43 Colorado tick fever segment 11 T RC 44 Colorado tick fever segment 12 U 45 Colorado tick fever segment 12 T 46 Colorado tick fever segment 12 U RC 47 Colorado tick fever segment 12 T RC 48 WO 2007/140506 PCT/AU2006/000755 62 Sequence Microorganism Sequence SEQ ID Listing # NO: 59 Foot and Mouth Foot and Mouth virus A U 1 Disease Foot and Mouth virus A T 2 Foot and Mouth virus A U RC 3 Foot and Mouth virus A T RC 4 Foot and Mouth virus Asia 1 U 5 Foot and Mouth virus Asia 1 T 6 Foot and Mouth virus Asia 1 U RC 7 Foot and Mouth virus Asia 1 T RC 8 Foot and Mouth virus C U 9 Foot and Mouth virus C T 10 Foot and Mouth virus C U RC 11 Foot and Mouth virus C T RC 12 Foot and Mouth virus 0 U 13 Foot and Mouth virus 0 T 14 Foot and Mouth virus 0 U RC 15 Foot and Mouth virus 0 T RC 16 Foot and Mouth virus SAT I1 U 17 Foot and Mouth virus SAT 1 T 18 Foot and Mouth virus SAT 1 U RC 19 Foot and Mouth virus SAT 1 T RC 20 Foot and Mouth virus SAT 2 U 21 Foot and Mouth virus SAT 2 T 22 Foot and Mouth virus SAT 2 U RC 23 Foot and Mouth virus SAT 2 T RC 24 Foot and Mouth virus SAT 3 U 25 Foot and Mouth virus SAT 3 T 26 Foot and Mouth virus SAT 3 U RC 27 Foot and Mouth virus SAT 3 T RC 28 60 Hepatitis GB virus GB virus C U 1 GB virus CT 2 GB virus C U RC 3 GB virus CT RC 4 61 Hendra virus Hendra virus U 1 Hendra virus T 2 Hendra virus U RC 3 Hendra virus T RC 4 62 Human adenovirus Human adenovirus A U 1 Human adenovirus A T 2 Human adenovirus A U RC 3 WO 2007/140506 PCT/AU2006/000755 63 Sequence Microorganism Sequence SEQ ID Listing # NO: Human adenovirus A T RC 4 Human adenovirus B U 5 Human adenovirus B T 6 Human adenovirus B U RC 7 Human adenovirus B T RC 8 Human adenovirus C U 9 Humanadenovirus CT 10 Human adenovirus C RC 11 Human adenovirus CT RC 12 Human adenovirus D U 13 Human adenovirus D T 14 Human adenovirus D U RC 15 Human adenovirus D T RC 16 Human adenovirus E U 17 Human adenovirus E T 18 Human adenovirus E U RC 19 Human adenovirus E T RC 20 Human adenovirus F U 21 Human adenovirus F T 22 Human adenovirus F U RC 23 Human adenovirus F T RC 24 63 Human astrovirus Human astrovirus U 1 Human astrovirus T 2 Human astrovirus U RC 3 Human astrovirus T RC 4 64 Human bocavirus Human bocavirus U 1 Human bocavirus T 2 Human bocavirus U RC 3 Human bocavirus T RC 4 65 Human coronavirus Human coronavirus 229E U 1 Human coronavirus 229E T 2 Human coronavirus 229E U RC 3 Human coronavirus 229E T RC 4 Human coronavirusHKU1 type A U 5 Human coronavirus HKU1 type A T 6 Human coronavirus HKU1 type A U RC 7 Human coronavirus HKU1 type A T RC 8 Human coronavirus NL63 U 9 Human coronavirus NL63 T 10 WO 2007/140506 PCT/AU2006/000755 64 Sequence Microorganism Sequence SEQ ID Listing # NO: Human coronavirus NL63 U RC 11 Human coronavirus NL63 T RC 12 Human coronavirus OU43 U 13 Human coronavirus OU43 T 14 Human coronavirus OU43 U RC 15 Human coronavirus OU43 T RC 16 66 Human enterovirus Human enterovirus-A U 1 Human enterovirus-A T 2 Human enterovirus A-U RC 3 Human enterovirus A-T RC 4 Human enterovirus-B U 5 Human enterovirus-B T 6 Human enterovirus-B U RC 7 Human enterovirus-B T RC 8 Human enterovirus-C U 9 Human enterovirus-C T 10 Human enterovirus-C U RC 11 Human enterovirus-C T RC 12 Human enterovirus-D U 13 Human enterovirus-D T 14 Human enterovirus-D U RC 15 Human enterovirus-D T RC 16 67 Human Herpes Virus Human Herpes Virus-1 U 1 Human Herpes Virus-1 T 2 Human Herpes Virus-1 U RC 3 Human Herpes Virus-1 T RC 4 Human Herpes Virus-2 U 5 Human Herpes Virus-2 T 6 Human Herpes Virus-2 U RC 7 Human Herpes Virus-2 T RC 8 Human Herpes Virus-3 U 9 Human Herpes Virus-3 T 10 Human Herpes Virus-3 U RC 11 Human Herpes Virus-3 T RC 12 Human Herpes Virus-4 U 13 Human Herpes Virus-4 T 14 Human Herpes Virus-4 U RC 15 Human Herpes Virus-4 T RC 16 Human Herpes Virus-5 (AD169) U 17 WO 2007/140506 PCT/AU2006/000755 65 Sequence Microorganism Sequence SEQ ID Listing # NO: Human Herpes Virus-5 (AD169) T 18 Human Herpes Virus-5 (AD169) U RC 19 Human Herpes Virus-5 T (AD169) RC 20 Human Herpes Virus-6A U 21 Human Herpes Virus-6A T 22 Human Herpes Virus-6A URC 23 Human Herpes Virus-6A T RC 24 Human Herpes Virus-6B U 25 Human Herpes Virus-6B T 26 Human Herpes Virus-6B URC 27 Human Herpes Virus-6B T RC 28 Human Herpes Virus-7 U 29 Human Herpes Virus-7 T 30 Human Herpes Virus-7 U RC 31 Human Herpes Virus-7 T RC 32 Human Herpes Virus-8 U 33 Human Herpes Virus-8 T 34 Human Herpes Virus-8 U RC 35 Human Herpes Virus-8 T RC 36 68 Human Human metapneumovirus U 1 metapneumovirus Human metapneumovirus T 2 Human metapneumovirus U RC 3 Human metapneumovirus T RC 4 69 Human Human Parainfluenza-1 U 1 Parainfluenza Human Parainfluenza-1 T 2 Human Parainfluenza-1 U RC 3 Human Parainfluenza-1 T RC 4 Human Parainfluenza-2 U 5 Human Parainfluenza-2 T 6 Human Parainfluenza-2 U RC 7 Human Parainfluenza-2 T RC 8 Human Parainfluenza-3 U 9 Human Parainfluenza-3 T 10 Human Parainfluenza-3 U RC 11 Human Parainfluenza-3 T RC 12 70 Human parechovirus Human parechovirus U 1 Human parechovirus T 2 Human parechovirus U RC 3 SHuman parechovirus T RC 4 WO 2007/140506 PCT/AU2006/000755 66 Sequence Microorganism Sequence SEQ ID Listing # NO: 71 Human Rhinovirus Human Rhinovirus-A U 1 Human Rhinovirus-A T 2 Human Rhinovirus-A U RC 3 Human Rhinovirus-A T RC 4 Human Rhinovirus-B U 5 Human Rhinovirus-B T 6 Human Rhinovirus-B U RC 7 Human Rhinovirus-B T RC 8 72 Human respiratory Human RSV U 1 syncytial virus Human RSV T 2 Human RSV U RC 3 Human RSV T RC 4 73 Measles virus Measles virus U 1 Measles virus T 2 Measles virus RC U 3 Measles virus RC T 4 74 Mumps virus Mumps virus U 1 Mumps virus T 2 Mumps virus RC U 3 Mumps virus RC T 4 75 Norovirus Norovirus U 1 Norovirus T 2 Norovirus RC U 3 Norovirus RC T 4 76 Norwalk virus Norwalk virus U 1 Norwalk virus T 2 Norwalk virus RC U 3 Norwalk virus RC T 4 77 Parvovirus B19 Parvovirus B19 U 1 Parvovirus B19 T 2 Parvovirus B19 RC U 3 Parvovirus B19 RC T 4 78 Poliovirus. Poliovirus U 1 Poliovirus T 2 Poliovirus RC U 3 Poliovirus RC T 4 79 Rabies virus Rabies virus U 1 Rabies virus T 2 Rabies virus RC U 3 WO 2007/140506 PCT/AU2006/000755 67 Sequence Microorganism Sequence SEQ ID Listing # NO: Rabies virus RC T 4 80 Ross River virus Ross River virus U 1 Ross River virus T 2 Ross River virus RC U 3 Ross River virus RC T 4 81 Rotavirus Rotavirus A NSP1 U 1 Rotavirus A NSP1 T 2 Rotavirus A NSP1 U RC 3 Rotavirus A NSP1 T RC 4 Rotavirus B NSP1 U 5 Rotavirus B NSP1 T 6 Rotavirus B NSP1 U RC 7 Rotavirus B NSP1 T RC 8 Rotavirus C NSP1 U 9 Rotavirus C NSP1 T 10 Rotavirus C NSP1 U RC 11 Rotavirus C NSP1 T RC 12 Rotavirus A NSP2 U 13 Rotavirus A NSP2 T 14 Rotavirus A NSP2 U RC 15 Rotavirus A NSP2 T RC 16 Rotavirus B NSP2 U 17 Rotavirus B NSP2 T 18 Rotavirus B NSP2 U RC 19 Rotavirus B NSP2 T RC 20 Rotavirus C NSP2 U 21 Rotavirus C NSP2 T 22 Rotavirus C NSP2 U RC 23 Rotavirus C NSP2 T RC 24 Rotavirus A NSP3 U 25 Rotavirus A NSP3 T 26 Rotavirus A NSP3 U RC 27 Rotavirus.A NSP3 T RC 28 Rotavirus B NSP3 U 29 Rotavirus B NSP3 T 30 Rotavirus B NSP3 U RC 31 Rotavirus B NSP3 T RC 32 Rotavirus C NSP3 U 33 Rotavirus C NSP3 T 34 WO 2007/140506 PCT/AU2006/000755 68 Sequence Microorganism Sequence SEQ ID Listing # NO: Rotavirus C NSP3 U RC 35 Rotavirus C NSP3 T RC 36 Rotavirus A NSP4 U 37 Rotavirus A NSP4 T 38 Rotavirus A NSP4 U RC, 39 Rotavirus A NSP4 T RC 40 Rotavirus B NSP4-U 41 Rotavirus B NSP4 T 42 Rotavirus B NSP4 U RC 43 Rotavirus B NSP4 T RC 44 Rotavirus C NSP4 U 45 Rotavirus C NSP4 T 46 Rotavirus C NSP4 U RC 47 Rotavirus C NSP4 T RC 48 Rotavirus A NSP5 U 49 Rotavirus A NSP5 T 50 Rotavirus A NSP5 U RC 51 Rotavirus A NSP5 T RC 52 Rotavirus B NSP5 U 53 Rotavirus B NSP5 T 54 Rotavirus B NSP5 U RC 55 Rotavirus B NSP5 T RC 56 Rotavirus C NSP5 U 57 Rotavirus C NSP5 T 58 Rotavirus C NSP5 U RC 59 Rotavirus C NSP5 T RC 60 Rotavirus A VP1 U 61 Rotavirus A VP1 T 62 Rotavirus A VP1 U RC 63 Rotavirus A VP1 T RC 64 Rotavirus C VP1 U 65 Rotavirus C VP1 T 66 Rotavirus C VP1 U RC 67 Rotavirus C VP1 T RC 68 Rotavirus A VP2 U 69 Rotavirus A VP2 T 70 Rotavirus A VP2 U RC 71 Rotavirus A VP2 T RC 72 Rotavirus B VP2 U 73 WO 2007/140506 PCT/AU2006/000755 69 Sequence Microorganism Sequence SEQ ID Listing # NO: Rotavirus B VP2 T 74 Rotavirus B VP2 U RC 75 Rotavirus B VP2 T RC 76 Rotavirus C VP2 U 77 Rotavirus C VP2 T 78 Rotavirus C VP2 U RC 79 Rotavirus C VP2 T RC 80 Rotavirus A VP3 U 81 Rotavirus A VP3 T 82 Rotavirus A VP3 U RC 83 Rotavirus A VP3 T RC 84 Rotavirus C VP3 U 85 Rotavirus C VP3 T 86 Rotavirus C VP3 U RC 87 Rotavirus C VP3 T RC 88 Rotavirus A VP4 U 89 Rotavirus A VP4 T 90 Rotavirus A VP4 U RC 91 Rotavirus A VP4 T RC 92 Rotavirus B VP4 U 93 Rotavirus B VP4 T 94 Rotavirus B VP4 U RC 95 Rotavirus B VP4 T RC 96 Rotavirus C VP4 U 97 Rotavirus C VP4 T 98 Rotavirus C VP4 U RC 99 Rotavirus C VP4 T RC 100 Rotavirus A VP6 U 101 Rotavirus A VP6 T 102 Rotavirus A VP6 U RC 103 Rotavirus A VP6 T RC 104 Rotavirus B VP6 U 105 Rotavirus B VP6 T 106 Rotavirus B VP6 U RC 107 Rotavirus B VP6 T RC 108 Rotavirus C VP6 U 109 Rotavirus C VP6 T 110 Rotavirus C VP6 U RC 111 Rotavirus C VP6 T RC 112 WO 2007/140506 PCT/AU2006/000755 70 Sequence Microorganism Sequence SEQ ID Listing # NO: Rotavirus A VP7 U 113 Rotavirus A VP7 T 114 Rotavirus A VP7 U RC 115 Rotavirus A VP7 T RC 116 Rotavirus B VP7 U 117 Rotavirus B VP7 T 118 Rotavirus B VP7 U RC 119 Rotavirus B VP7 T RC 120 Rotavirus C VP7 U 121 Rotavirus C VP7 T 122 Rotavirus C VP7 U RC 123 Rotavirus C VP7 T RC 124 82 SARS coronavirus SARS coronavirus U 1 SARS coronavirus T 2 SARS coronavirus RC U 3 SARS coronavirus RC T 4 83 TT virus TT virus U 1 TT virus T 2 TT virus RC U 3 TT virus RC T 4 84 TTV minivirus TTV minivirus U 1 TTV minivirus T 2 TTV minivirus U RC 3 TTV minivirus T RC 4 85 West Nile virus West Nile virus U 1 West Nile virus T 2 West Nile virus U RC 3 West Nile virus T RC 4 86 Alpha Virus Alpha Virus U 1 Alpha Virus T 2 Alpha Virus U RC 3 Alpha Virus T RC 4 87 Camel pox Camel pox U 1 Camel pox T 2 Camel pox U RC 3 Camel pox T RC 4 88 Cow pox Cow pox U 1 Cowpox T 2 Cow pox U RC 3 WO 2007/140506 PCT/AU2006/000755 71 Sequence Microorganism Sequence SEQ ID Listing # NO: Cow pox T RC 4 89 Coxiella burnetii Coxiella burnetii U 1 Coxiella burnetil T 2 Coxiella burnetii U RC 3 Coxiella burnetii T RC 4 90 Crimean-Congo HF Crimean-Congo HF S U 1 Crimean-Congo HF S T 2 Crimean-Congo HF S U RC 3 Crimean-Congo HF S T RC 4 Crimean-Congo HF L U 5 Crimean-Congo HF L T 6 Crimean-Congo HF L U RC 7 Crimean-Congo HF L T RC 8 Crimean-Congo HF M U 9 Crimean-Congo HF M T 10 Crimean-Congo HF M U RC 11 Crimean-Congo HF M T RC 12 91 Dengue virus Dengue virus-1 U 1 Dengue virus-1 T 2 Dengue virus-1 U RC 3 Dengue virus-1 T RC 4 Dengue virus-2 U 5 Dengue virus-2 T 6 Dengue virus-2 U RC 7 Dengue virus-2 T RC 8 Dengue virus-3 U 9 Dengue virus-3 T 10 Dengue virus-3 U RC 11 Dengue virus-3 T RC 12 Dengue virus-4 U 13 Dengue virus-4 T 14 Dengue virus-4 U RC 15 Dengue virus-4 T RC 16 92 Eastern Equine Eastern Equine Encephalitis U 1 Encephalitis Eastern Equine Encephalitis T 2 Eastern Equine Encephalitis U RC 3 Eastern Equine Encephalitis T RC 4 93 Ebola virus Ebola Sudan U 1 WO 2007/140506 PCT/AU2006/000755 72 Sequence Microorganism Sequence SEQ ID Listing # NO: Ebola Sudan T 2 Ebola Sudan U RC 3 Ebola Sudan T RC 4 Ebola Reston U 5 Ebola Reston T 6 Ebola Reston U RC 7 Ebola Reston T RC 8 Ebola Zaire U 9 Ebola Zaire T 10 Ebola Zaire U RC 11 Ebola Zaire T RC 12 94 Marburg virus Marburg virus U 1 Marburg virus T 2 Marburg virus U RC 3 Marburg virus T RC 4 95 Guanarito virus Guanarito virus L U 1 Guanarito virus L T 2 Guanarito virus L U RC 3 Guanarito virus L T RC 4 Guanarito virus S U 5 Guanarito virus S T 6 Guanarito virus S U RC 7 Guanarito virus S T RC 8 96 Hanta virus Hanta virus M U 1 Hanta virus MT 2 Hanta virus M U RC 3 Hanta virus MT RC 4 Hanta virus L U 5 Hanta virus L T 6 Hanta virus L U RC 7 Hanta virus L T RC 8 Hanta virus S U 9 Hanta virus ST 10 Hanta virus S U RC 11 Hanta virus ST RC 12 97 Hantaan virus Hantaan virus L U 1 Hantaan virus L T 2 Hantaan virus LU RC 3 Hantaan virus L T RC 4 WO 2007/140506 PCT/AU2006/000755 73 Sequence Microorganism Sequence SEQ ID Listing # NO: Hantaan virus M U 5 Hantaan virus MT 6 Hantaan virus L M RC 7 Hantaan virus L M RC 8 Hantaan virus S U 9 Hantaan virus ST 10 Hantaan virus S U RC 11 Hantaan virus S T RC 12 98 Japanese Japanese encephalitis virus U 1 encephalitis virus Japanese encephalitis virus T 2 Japanese encephalitis virus U RC 3 Japanese encephalitis virus T RC 4 99 Junin virus Junin virus S U 1 Junin virus ST 2 Junin virus S U RC 3 Junin virus ST RC 4 Junin virus LU 5 Junin virus L T 6 Junin virus LU RC 7 Junin virus L T RC 8 100 Lassa virus Lassa virus LU 1 Lassa virus L T 2 Lassa virus LU RC 3 Lassa virus L T RC 4 Lassa virus S U 5 Lassa virus ST 6 Lassa virus S U RC 7 Lassa virus ST RC 8 101 Machupo virus Machupo virus L U 1 Machupo virus L T 2 Machupo virus L U RC 3 Machupo virus L T RC 4 Machupo virus S U 5 Machupo virus ST 6 Machupo virus S U RC 7 Machupo virus S T RC 8 102 Monkeypox virus Monkeypox virus U 1 Monkeypox virus T 2 WO 2007/140506 PCT/AU2006/000755 74 Sequence Microorganism Sequence SEQ ID Listing # NO: Monkeypox virus U RC 3 Monkeypox virus T RC 4 103 Murray Valley Murray Valley encephalitis U 1 encephalitis Murray Valley encephalitis T 2 Murray Valley encephalitis U RC 3 Murray Valley encephalitis T RC 4 104 Nipah virus Nipah virus U 1 Nipah virus T 2 Nipah virus U RC 3 Nipah virus T RC 4 105 Rift Valley Fever Rift Valley Fever virus S U 1 virus Rift Valley Fever virus S T 2 Rift Valley Fever virus SU RC 3 Rift Valley Fever virus S T RC 4 Rift Valley Fever virus M U 5 Rift Valley Fever virus MT 6 Rift Valley Fever virus MU RC 7 Rift Valley Fever virus MT RC 8 Rift Valley Fever virus L U 9 Rift Valley Fever virus LT 10 Rift Valley Fever virus LU RC 11 Rift Valley Fever virus L T RC 12 106 Sabia virus Sabia virus S U 1 Sabia virus ST 2 Sabia virus S U RC 3 Sabia virus ST RC 4 Sabia virus LU 5 Sabia virus L T 6 Sabia virus LU RC 7 Sabia virus L T RC 8 107 Sin Nombre virus Sin Nombre virus L U 1 Sin Nombre virus L T 2 Sin Nombre virus L U RC 3 Sin Nombre virus L T RC 4 Sin Nombre virus S U 5 Sin Nombre virus ST 6 Sin Nombre virus S U RC 7 Sin Nombre virus S T RC 8 Sin Nombre virus M U 9 WO 2007/140506 PCT/AU2006/000755 75 Sequence Microorganism Sequence SEQ ID Listing # NO: Sin Nombre virus M T 10 Sin Nombre virus M U RC 11 Sin Nombre virus M T RC 12 108 Variola major Variola major U 1 Variola major T 2 Variola major U RC 3 Variola major T RC 4 109 Variola minor Variola minor U 1 Variola minor T 2 Variola minor U RC 3 Variola minor T RC 4 110 Venezuelan equine Venezuelan equine encephalitis U 1 encephalitis Venezuelan equine encephalitis T 2 Venezuelan equine encephalitis U RC 3 Venezuelan equine encephalitis T RC 4 111 Western equine Western equine encephalitis U 1 encephalitis Western equine encephalitis T 2 Western equine encephalitis U RC 3 Western equine encephalitis T RC 4 112 Yellow Fever virus Yellow Fever virus U 1 Yellow Feyer virus T 2 Yellow Fever virus U RC 3 Yellow Fever virus T RC 4 *U derivative top strand with all cytosine replaced with uracil *T modified top strand with all uracil replaced with thymine *U RC derivative bottom strand with all cytosine replaced with uracil *T RC modified bottom strand with all uracil replaced with thymine 5 It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and 10 not restrictive.

Claims (86)

1. A derivative or modified nucleic acid for Hepatitis C virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 76 in Sequence Listing #51, parts thereof comprising at least about 20 nucleotides, and nucleic acid 5 molecules capable of hybridizing thereto under stringent conditions.
2. A derivative or modified nucleic acid for Acinetobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #1, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 10 3. A derivative or modified nucleic acid for Bacillus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #2, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
4. A derivative or modified nucleic acid for Bacteroides sp having a sequence 15 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #3, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
5. A derivative or modified nucleic acid for Bartonella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence 20 Listing #4, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
6. A derivative or modified nucleic acid for Bordetella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #5, parts thereof comprising at least about 20 nucleotides, and nucleic acid 25 molecules capable of hybridizing thereto under stringent conditions.
7. A derivative or modified nucleic acid for Borrelia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #6, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 30 8. A derivative or modified nucleic acid for Brucella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #7, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 77
9. A derivative or modified nucleic acid for Campylobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #8, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 10. A derivative or modified nucleic acid for Chlamydia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #9, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
11. A derivative or modified nucleic acid for Clostridium sp having a sequence 10 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 39 in Sequence Listing #10, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
12. A derivative or modified nucleic acid for Cornebacterium sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence 15 Listing #11, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
13. A derivative or modified nucleic acid for Escherichia col having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #12, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
14. A derivative or modified nucleic acid for Ehrlichia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #13, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 25 15. A derivative or modified nucleic acid for Enterococcus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #14, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
16. A derivative or modified nucleic acid for Fusobacterium sp having a sequence 30 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #156, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
17. A derivative or modified nucleic acid for Haemophilus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence WO 2007/140506 PCT/AU2006/000755 78 Listing #16, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
18. A derivative or modified nucleic acid for Helicobacter sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 5 Listing #17, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
19. A derivative or modified nucleic acid for Legionella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #18, parts thereof comprising at least about 20 nucleotides, and nucleic acid 10 molecules capable of hybridizing thereto under stringent conditions.
20. A derivative or modified nucleic acid for Leptospira sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #19, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 15 21. A derivative or modified nucleic acid for Listeria sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #20, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
22. A derivative or modified nucleic acid for Mycobacterium sp having a sequence 20 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #21, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
23. A derivative or modified nucleic acid for Mycoplasma sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 25 Listing #22, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
24. A derivative or modified nucleic acid for Neisseria sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #23, parts thereof comprising at least about 20 nucleotides, and nucleic acid 30 molecules capable of hybridizing thereto under stringent conditions.
25. A derivative or modified nucleic acid for Norcadia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #24, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 79
26. A derivative or modified nucleic acid for Pseudomonas sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #25, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 27. A derivative or modified nucleic acid for Rickettsia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #26, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
28. A derivative or modified nucleic acid for Salmonella sp having a sequence 10 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #27, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
29. A derivative or modified nucleic acid for Seratia sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing 15 #28, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
30. A derivative or modified nucleic acid for Shigella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #29, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
31. A derivative or modified nucleic acid for Staphylococcus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 52 in Sequence Listing #30, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 25 32. A derivative or modified nucleic acid for Streptococcus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #31, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
33. A derivative or modified nucleic acid for Streptomyces having a sequence 30 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #32, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
34. A derivative or modified nucleic acid for Treponema sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence WO 2007/140506 PCT/AU2006/000755 80 Listing #33, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
35. A derivative or modified nucleic acid for Trophermya sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 5 Listing #34, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
36. A derivative or modified nucleic acid for Plasmodium sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO:.60 in Sequence Listing #35, parts thereof comprising at least about 20 nucleotides, and nucleic acid 10 molecules capable of hybridizing thereto under stringent conditions.
37. A derivative or modified nucleic acid for Aspergillis sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #36, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 15 38. A derivative or modified nucleic acid for Candida sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #37, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
39. A derivative or modified nucleic acid for Cryptococcus sp having a sequence 20 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #38, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
40. A derivative or modified nucleic acid for Paracoccidioides sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 25 Listing #39, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
41. A derivative or modified nucleic acid for Rhizopus sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #40, parts thereof comprising at least about 20 nucleotides, and nucleic acid 30 molecules capable of hybridizing thereto under stringent conditions. .42. A derivative or modified nucleic acid for Francisella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #41, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 81
43. A derivative or modified nucleic acid for Vibrio sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #42, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 44. A derivative or modified nucleic acid for Yersinia sp having a sequence selected. from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #43, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
45. A derivative or modified nucleic acid for JC polyomavirus having a sequence 10 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #44, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
46. A derivative or modified nucleic acid for Andes virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing 15 #46, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
47. A derivative or modified nucleic acid for hepatitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #52, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
48. A derivative or modified nucleic acid for Human Immunodeficiency virus (HIV) having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 128 in Sequence Listing #53, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under 25 stringent conditions.
49. A derivative or modified nucleic acid for Influenza virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 162 in Sequence Listing #54, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 30 50. A derivative or modified nucleic acid for BK virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #55, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 82
51. A derivative or modified nucleic acid for Barmah virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #56, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 52. A derivative or modified nucleic acid for Calcivirus virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #57, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
53. A derivative or modified nucleic acid for Colorado tick fever virus having a 10 sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 48 in Sequence Listing #58, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
54. A derivative or modified nucleic acid for Foot and Mouth virus having a sequence selected from the group consisting of SEQ ID NO: I1 to SEQ ID NO: 28 in Sequence 15 Listing #59, parts thereof comprising at least about 20 nucleotides, and nucleic acid Molecules capable of hybridizing thereto under stringent conditions.
55. A derivative or modified nucleic acid for Hepatitis GB virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #60, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
56. A derivative or modified nucleic acid for Henda virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #61, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 25 57. A derivative or modified nucleic acid for Human adenovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 24 in Sequence Listing #62, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hyLridizing thereto under stringent conditions.
58. A derivative or modified nucleic acid for Human astrovirus having a sequence 30 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #63, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
59. A derivative or modified nucleic acid for Human bocavirus virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in WO 2007/140506 PCT/AU2006/000755 83 Sequence Listing #64, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
60. A derivative or modified nucleic acid for Human coronavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence 5 Listing #65, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
61. A derivative or modified nucleic acid for Human enterovirus virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #66, parts thereof comprising at least about 20 nucleotides, and 10 nucleic acid molecules capable of hybridizing thereto under stringent conditions.
62. A derivative or modified nucleic acid for Human herpes virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 36 in Sequence Listing #67, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 15 63. A derivative or modified nucleic acid for Human metapneumovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in )Sequence Listing #68, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
64. A derivative or modified nucleic acid for Human parainfluenzavirus having a 20 sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #69, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
65. A derivative or modified nucleic acid for Human parechovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 25 Listing #70, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
66. A derivative or modified nucleic acid for Human rhinovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #71, parts thereof comprising at least about 20 nucleotides, and nucleic acid 30 molecules capable of hybridizing thereto under stringent conditions.
67. A derivative or modified nucleic acid for Human respiratory syncytial virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #72, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 84
68. A derivative or modified nucleic acid for Measles virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #73, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 69. A derivative or modified nucleic acid for Mumps virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #74, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
70. A derivative or modified nucleic acid for Norovirus having a sequence selected 10 from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #75, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
71. A derivative or modified nucleic acid for Norwalk virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 15 Listing #76, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
72. A derivative or modified nucleic acid for Parvovirus B19having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #77, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
73. A derivative or modified nucleic acid for Poliovirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #78, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 25 74.. A derivative or modified nucleic acid for Rabies virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #79, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
75. A derivative or modified nucleic acid for Ross River virus having a sequence 30 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #80, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
76. A derivative or modified nucleic acid for Rotavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 124 in Sequence Listing WO 2007/140506 PCT/AU2006/000755 85 #81, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
77. A derivative or modified nucleic acid for SARS coronavirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 5 Listing #82, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
78. A derivative or modified nucleic acid for TT virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #83, parts thereof comprising at least about 20 nucleotides, and nucleic acid 10 molecules capable of hybridizing thereto under stringent conditions.
79. A derivative or modified nucleic acid for TTV minivirus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #84, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 15 80. A derivative or modified nucleic acid for West Nile virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #85, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
81. A derivative or modified nucleic acid for Alpha virus having a sequence selected 20 from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #86, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
82. A derivative or modified nucleic acid for Camel pox virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence 25 Listing #87, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
83. A derivative or modified nucleic acid for Cow Pox virus having a sequence selected from the group. consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #88, parts thereof comprising at least about 20 nucleotides, and nucleic acid 30 molecules capable of hybridizing thereto under stringent conditions.
84. A derivative or modified nucleic acid for Coxiella sp having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #89, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 86
85. A derivative or modified nucleic acid for Crimean-Congo HF having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #90, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 86. A derivative or modified nucleic acid for Dengue virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 16 in Sequence Listing #91, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
87. A derivative or modified nucleic acid for Eastern Equine Encephalitis virus having 10 a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #92, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
88. A derivative or modified nucleic acid for Ebola virus having a sequence selected from the group consisting ofSEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing 15 #93, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
89. A derivative or modified nucleic acid for Marburg virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #94, parts thereof comprising at least about 20 nucleotides, and nucleic acid 20 molecules capable of hybridizing thereto under stringent conditions.
90. A derivative or modified nucleic acid for Guanarito virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #95, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 25 91. A derivative or modified nucleic acid for Hanta virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #96, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
92. A derivative or modified nucleic acid for Hantan virus having a sequence selected 30 from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #97, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
93. A derivative or modified nucleic acid for Japanese encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in WO 2007/140506 PCT/AU2006/000755 87 Sequence Listing #97, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
94. A derivative or modified nucleic acid for Junin virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing 5 #99, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
95. A derivative or modified nucleic acid for Lassa virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #100, parts thereof comprising at least about 20 nucleotides, and nucleic acid 10 molecules capable of hybridizing thereto under stringent conditions.
96. A derivative or modified nucleic acid for Machupo virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #101, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 15 97. A derivative or modified nucleic acid for Monkey pox virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #102, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
98. A derivative or modified nucleic acid for Murray Valley encephalitis virus having a 20 sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #103, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
99. A derivative or modified nucleic acid for Nipah virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing 25 #104, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
100. A derivative or modified nucleic acid for Rift Valley Fever virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #105, parts thereof comprising at least about 20 nucleotides, and nucleic acid 30 molecules capable of hybridizing thereto under stringent conditions.
101. A derivative or modified nucleic acid for Sabia virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 8 in Sequence Listing #106, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. WO 2007/140506 PCT/AU2006/000755 88
102. A derivative or modified nucleic acid for Sin Nombre virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12 in Sequence Listing #107, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions. 5 103. A derivative or modified nucleic acid for Variola major virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #108, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
104. A derivative or modified nucleic acid for Variola minor virus having a sequence 10 selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #109, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
105. A derivative or modified nucleic acid for Venezuelan equine encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to 15 SEQ ID NO: 4 in Sequence Listing #110, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
106. A derivative or modified nucleic acid for Western equine encephalitis virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in 20 Sequence Listing #111, parts thereof comprising at least about 20 nucleotides, and nucleic acid molecules capable of hybridizing thereto under stringent conditions.
107. A derivative or modified nucleic acid for Yellow Fever virus having a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 in Sequence Listing #112, parts thereof comprising at least about 20 nucleotides, and nucleic acid 25 molecules capable of hybridizing thereto under stringent conditions.
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CA2680426A1 (en) * 2007-03-16 2008-09-25 Human Genetic Signatures Pty Ltd Assay for gene expression
FR2940805B1 (en) 2009-01-05 2015-10-16 Biomerieux Sa METHOD FOR AMPLIFYING AND / OR DETECTING NUCLEIC ACIDS, KITS AND USES THEREOF
WO2010121308A1 (en) * 2009-04-23 2010-10-28 Human Genetic Signatures Pty Ltd Detection of hepatitis c virus
US8628947B2 (en) 2010-08-30 2014-01-14 Intervet Inc. Potomac horse fever isolates
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US9775894B2 (en) 2013-07-09 2017-10-03 University Of Washington Through Its Center For Commercialization Methods and compositions for activation of innate immune responses through RIG-I like receptor signaling
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