JP2002355083A - Method for detecting nucleic acid related to disease - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method by which the objective information about a nucleic acid can simply and accurately be recovered from a sample substance in a short time. SOLUTION: This method is to obtain first information about the nucleic acid from an individual exposed to a specific disease and second information about a nucleic acid from a pathogenic microorganism present in the individual and the pathogenic microorganism is related to the specific disease and has the following. (a) reacting an extract of the nucleic acid from the individual with a probe immobilizing substrate having a first probe and a second probe wherein the first probe detects the presence of a specific nucleic acid sequence of the pathogenic microorganism, the pathogenic microorganism is related to the specific disease and the second probe detects the presence of the specific nucleic acid in the individual and (b) obtaining the first information by, if present, detecting the presence of the nucleic acid ligated with the first probe and obtaining the second information by, if present, detecting the presence of the nucleic acid ligated with the second probe.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a method for obtaining information on a nucleic acid associated with a disease obtained from an individual. The present invention relates to a method for obtaining information on the nucleic acid of an individual, as well as information on the nucleic acid obtained with respect to a disease, particularly information on the nucleic acid of a pathogenic microorganism associated with the disease.

[0002] The present invention also relates to a probe-immobilized substrate, particularly to a probe-immobilized chip for use in the above method.

[0003]

2. Description of the Related Art Genetic testing techniques using DNA chips have attracted attention (Beattie et al., Clin Chem 39: 719-22, Fodor et al.
al., Science, 251, 767 (1991), Khrapko et al., FE
BS Lett, 256, 118 (1989), and Southern et al., Nuc
leic Acids Res., 22, 1368 (1994)). The DNA chip is a glass or silicon chip of several cm square in which a plurality of types of DNA probes having different sequences are immobilized. The DNA probe on such a chip is reacted with a sample nucleic acid labeled with a fluorescent dye or a radioisotope (RI), or a mixture of an unlabeled sample nucleic acid and a labeled oligonucleotide. When a sequence complementary to the DNA probe immobilized on the chip is present in the sample, a signal derived from the label used at a specific site on the chip is obtained. Therefore, if the sequence (sequence) and position of the immobilized DNA probe are known in advance, the base sequence (sequen
ce) (Pease et al., Proc. Nat
l. Acad. Sci. USA, 91, 5022 (1994), Parinov et al.,
Nucleic Acids Res., 24, 2998 (1996). .

[0004] By obtaining a sample substance such as blood obtained from a patient and extracting a nucleic acid from the sample substance, a nucleic acid unique to the patient can be obtained. If the patient-specific nucleic acid is further subjected to genetic analysis, the susceptibility to a certain disease,
It is possible to obtain information related to the patient's constitution, such as the efficacy of the drug and / or the likelihood of side effects. As a result, it is possible to obtain information specific to the patient. Therapies designed specifically for individual patients are called "tailor-made medicine."

[0005] In carrying out tailor-made medical treatment, there is a demand for the development of a method for predicting a treatment method for a disease more accurately and simply from a sample substance extracted from a patient.

[0006]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method capable of easily and accurately recovering target nucleic acid information from a sample substance in a short time.

[0007]

Means for solving the above-mentioned problems include first information on nucleic acid of an individual exposed to a specific disease and second information on nucleic acid from a pathogenic microorganism present in the individual. Wherein the pathogenic microorganism is associated with the specific disease and comprises: (a) an extract of a nucleic acid from the individual, a first probe and a second probe; Reacting with a probe-immobilized substrate comprising: wherein the first probe detects the presence of a specific nucleic acid sequence of the pathogenic microorganism, wherein the pathogenic microorganism is associated with the specific disease, and The second probe detects the presence of a specific nucleic acid of the individual; and (b) detecting the presence of a nucleic acid bound to the first probe as a result of the reaction of (a). Get 1 information And obtaining the second information by detecting the presence of the nucleic acid bound to the second probe: and detecting the presence of the substrate and the specific nucleic acid sequence of the pathogenic microorganism. A first probe immobilized on a substrate, wherein the microorganism is different from the first probe and is immobilized on the substrate for detecting the presence of a specific nucleic acid in an individual associated with a specific disease; A probe-immobilized substrate comprising a second probe having a sequence, wherein the nucleic acid of the individual is involved in responsiveness to treatment of the disease.

[0008]

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for obtaining information about nucleic acids from an individual exposed to a disease. The individual may be at an early or late stage of the disease. The individual may be an individual suspected of suffering from the disease or an individual suspected of having the disease in the future.

The present invention relates to the correlation of nucleic acid information from both the individual and the disease. In embodiments disclosed herein, the individual has been exposed to a disease associated with a pathogenic microorganism such as, for example, a virus, bacteria, yeast, or mycoplasma. The present invention also relates to a correlation between nucleic acid information obtained from pathogenic microorganisms present in the individual and nucleic acids from the individual.

[0010] Thus, in one aspect, the present invention provides information about nucleic acids associated with a particular disease in an individual, particularly information about nucleic acids associated with the susceptibility of said individual to treatment for a disease, and the presence of said individual in said individual. A method is also provided for obtaining information on nucleic acids associated with the specific disease derived from a pathogenic microorganism.

[0011] The present invention also relates to a specific nucleic acid probe used in identifying a nucleic acid from the individual and a nucleic acid from the pathogenic microorganism.

[0012] In some embodiments, the nucleic acid probe is immobilized on a substrate such as a chip. The present invention also relates to a probe-immobilized substrate, for example, a probe-immobilized chip for use in the method.
Other substrates, for example, DNA capillary electrophoresis devices, beads, etc., membrane-based arrays, microtiter plates, electrodes, semiconductor-based devices, etc., waveguide materials, surface plasmon resonance (Surface pla
smon resonance; hereinafter referred to as SPR), Quartz crystal microbalance;
QCM) and mass spectroscopy can also be used in the present invention. Furthermore, allele-specific oligo hybridization, restriction method (microtiterarray diagonal gel electrophoresis; microtitierarray
diagonal gel electrophoresis), ligation method (padlock probe, oligonucleotide ligation assay, dye-labeled oligonucleotide ligation), nucleotide incorporation (mini-sequencing, template-directed dye-termination assay (template-
Directed dye-termination assay)) and invader assay can also be used in the present invention. Other common solution hybridization techniques can also be used in the present invention, for example, amplification methods (polymerase chain reaction; PCR),
Ligase chain reaction (LCR), nucleic acid sequence-based amplification (nu
cleic acid sequence-based amplification (NASBA),
Strand displacement amplification (strand displac)
ement amplification (SDA), loop-media
Isothermal amplification (loop-mediated isothermal amplifi)
cation; LAMP), isothermal chimeric primer initiation amplification of nucleic acid
(isothermal and chimeric primer-initiated amplific
ICAN), branched DNA that can be used to detect the solid and the nucleic acid of the pathogenic microorganism, and the like can be used.

The sample substance containing nucleic acids collected from the individual may be whole blood, blood, serum, leukocytes, urine, stool, semen, saliva, biopsy tissue, cultured cells, sputum, or the like. The preferred sample is blood. Also, the sample substance is a nucleic acid derived from the individual associated with susceptibility to treatment for a disease,
It is preferable that the nucleic acid contains the nucleic acid associated with the pathogenic microorganism present in the individual. Here, the pathogenic microorganism is associated with the disease. Further, the sample may have been subjected to any necessary pretreatment such as homogenate and extraction, if necessary. Such a pretreatment could be selected by one skilled in the art depending on the biological sample of interest.

In the illustrative embodiment disclosed herein, a whole blood sample is first collected from a human. Next, from the collected whole blood sample, for example, QIAamp DNA
Blood Midi Kid (registered trademark, QIAG)
Using a commercially available kit for human genome extraction such as EN, Tokyo, Japan), nucleic acids derived from the human genome and nucleic acids derived from the virus genome present in the human are both extracted. In some embodiments, the nucleic acid containing the sequence of interest is amplified. continue,
The obtained amplification product is hybridized to the probe-immobilized chip, and the resulting amplified product is analyzed by detecting a nucleic acid that binds to the probe-immobilized chip. In this way, information on the nucleic acid associated with the specific disease from the individual is obtained along with information on the nucleic acid associated with the specific disease obtained from the pathogenic microorganisms present in the individual.

In the above method, an example of a human individual has been described, but the individual is not limited to a human. According to aspects of the present invention, an example of an "individual" of interest may be any mammal, including a human, dog, cat, cow, goat, pig, sheep, or monkey. However, humans are the most preferred individuals. Also, according to aspects of the invention, the individual may be a healthy individual or an individual suffering from some disease. However, in general, by performing on an individual suffering from a disease caused by the pathogenic microorganism and the type of nucleic acid possessed by the individual, the method according to the invention will be used more effectively. As used herein, the term "certain disease" includes diseases associated with pathogenic microorganisms, such as viruses, bacteria, yeasts or mycoplasmas, as well as oncological diseases, and preferably diseases associated with pathogenic microorganisms. More preferably, its onset and therapeutic effect depend on the type of nucleic acid contained in the individual suffering from the disease and / or the presence or absence of expression of such a nucleus-containing gene. It is a disease. However, the present invention is not particularly limited to these diseases.

As used herein, the term "target nucleic acid" refers to a nucleic acid containing a sequence to be detected and / or analyzed.

According to the aspect of the present invention, the nucleic acid derived from an individual to be extracted may be any nucleic acid derived from an individual, and may be DNA or RNA, or genomic DNA. The nucleic acid derived from the pathogenic microorganism to be extracted may be any nucleic acid derived from the target pathogenic microorganism.
Or RNA.

A method for extracting a nucleic acid component from a sample substance is as follows.
The means is not particularly limited, and a liquid-liquid extraction method such as a phenol-chloroform method or a solid-liquid extraction method using a carrier can also be used. In addition, although not limited thereto, for example, nucleic acid extraction method QIAamp
Commercially available kits such as (QIAGEN), genomic DNA purification kit (Promega), and Sumytest (Sumitomo Metals) can be used. Then, P
An amplification operation such as CR may or may not be performed.

When the extracted nucleic acid component is RNA,
HCV-RNA may be used directly as a sample nucleic acid,
After preparing cDNA from RNA using reverse transcriptase, it may be used as a sample nucleic acid. Also in this case, it may be performed without separating nucleic acids from individuals and nucleic acids from pathogenic microorganisms,
For example, in the method described above, QIAamp DNAB
After performing extraction using a load Midi Kid, reverse transcription may be performed, followed by PCR amplification.

The amplification carried out according to the embodiment of the present invention may be any means for amplifying nucleic acids known per se, and is preferably carried out by the polymerase chain reaction (hereinafter referred to as PCR). If the sample substance contains a sufficient amount of nucleic acid, amplification may be omitted.

The probe-immobilized substrate such as a probe-immobilized chip that can be used according to the embodiment of the present invention is a probe-immobilized chip obtained by immobilizing a first probe and a second probe having mutually different base sequences on a substrate. . The first probe is a probe for detecting the presence of a specific nucleic acid sequence derived from a pathogenic microorganism associated with a specific disease. The second probe is associated with the specific disease,
It is a probe for detecting the presence of a specific nucleic acid sequence derived from an individual.

With such a probe-immobilized substrate, analysis can be performed simultaneously by using the same substrate for nucleic acids obtained from two different sources: an individual and a pathogenic microorganism. Such a probe-immobilized chip is disclosed for the first time by the present applicant. According to the present invention, such a probe-immobilized substrate is also provided as one aspect of the present invention.

In the probe-immobilized chip according to the embodiment of the present invention, the first probe detects a nucleic acid sequence derived from a pathogenic microorganism contained in a sample substance such as whole blood from a human patient. Such a probe may have a base sequence corresponding to a chromosomal DNA or a gene (RNA) derived from a pathogenic microorganism that may be infected by the target individual (eg, a human patient), Or its cDNA or cRNA, chromosomal DNA, R
It may have a nucleotide sequence corresponding to NA, cDNA and cRNA fragments or their complementary sequences.

The amount of the pathogenic microorganism in the sample substance is also measured using the above-mentioned first probe, that is, a probe having a nucleotide sequence corresponding to a nucleic acid sequence derived from the pathogenic microorganism, a fragment thereof, or a complementary sequence thereof. It is possible to measure.

Further, it is desirable that the first probe is a probe that detects not only the presence of the pathogenic microorganism in the sample substance but also a gene mutation of the pathogenic microorganism.

The measurement of the nucleic acid (RNA) expressed by the pathogenic microorganism in the sample substance is also the first probe described above, ie, a probe having a base sequence corresponding to a nucleic acid sequence derived from the pathogenic microorganism, a fragment thereof, or a complementary sequence thereof. Can be measured using The nucleic acid strand in this case is RNA or cD
NA or cRNA can be detected or quantified.

In the case where the pathogenic microorganism is a virus,
In the embodiment, the specific gene is expressed in large amounts during the growth phase of the virus, so that measuring its expression pattern (quantity, quality) makes it possible to predict drug efficacy.

In the probe-immobilized chip according to the embodiment of the present invention, as the second probe, a chromosomal DNA sequence, a gene (RNA) or its cDNA or cRNA, a fragment thereof or a complementary sequence thereof, which the individual has, is involved in the disease. And a base sequence corresponding to The “nucleic acid sequence related to a disease” possessed by an individual is a nucleic acid sequence present in a chromosome in the cells of the individual, and is a nucleic acid sequence capable of predicting responsiveness to treatment. More specifically, the nucleic acid sequence is, for example, a nucleic acid sequence that predicts the reactivity of an individual to a disease, or determines the efficacy and / or the likelihood of side effects of a drug to be administered.

With the second probe, it is possible to detect the presence of a specific nucleic acid sequence in an individual, the expression level of a specific gene, the gene expression pattern in an individual, and the like.

The first probe or the second probe according to the present invention has at least 11 bases, preferably at least 1 base.
2 bases, more preferably at least 13, 14, 15,
16, 17, 18, 19, 20, 21, 22, 23, 2,
It is desirable to have a base sequence of 4, 25, 26, 27, 28, 29 and 30 bases in length. If the length of the base sequence to be immobilized on the substrate is excessively long, it becomes difficult to discriminate one base difference. On the other hand, if the length of the base sequence immobilized on the substrate is too short, it becomes difficult to determine the base sequence of the base sequence contained in the sample.

As the first probe or the second probe, ribonucleic acid (RNA), deoxyribonucleic acid (DN
A), peptide nucleic acids (PNA), methylphosphonate nucleic acids, oligonucleotides such as S-oligos, and cDN
A, nucleic acids such as polynucleotides such as cRNA can be used.

According to an embodiment of the present invention, the probe-immobilized substrate is a substrate, a porous body (Beattie et al., Clin. Chem., 4).
1, 700 (1995)), microtiter plate (Kawai et.
al., Anal.Biochem., 209, 63 (1993)), beads (Mirki
n et al., Nature, 382, 607 (1996), spherical, particulate, magnetic, magnetic beads (Miller et al., J. Magne
t. Magn. Mater., 225, 138 (2001), DNA capillary el.
ectrophoresis chip (Chou et al., Proc. Natl. Acd.
Sci., 96, 11 (1999), membrane-based array (Lemieu
x et al., Molecular Breeding, 4, 277 (1998), semiconductor-based devices, etc. (Thewes et al., 2002 IEE
E International Solid-State Circuits Conference, 35
0 (2002), waveguide materials (Piunno et al., Anal.
m. Acta, 288, 205 (1994)), SPR (Nelson et al., Anal.
Chem., 73, 1 (2001), QCM (Ito et al., Anal.
cta, 327, 29 (1996), mass spectrometry (Amexis et al., Proc.
Natl. Acd. Sci., 98, 12097 (2001) and the like can be prepared by immobilizing a first probe and a second probe consisting of the above-mentioned nucleotide sequence on a substrate formed.
The material, size, shape and the like of the substrate on which the nucleic acid is to be immobilized are not particularly limited, and any substrate capable of immobilizing the nucleic acid may be used. Detection of the sequence of the nucleic acid in the sample substance using the probe-immobilized substrate, for example,
After extracting a nucleic acid component from a sample substance collected from an individual, the sample nucleic acid is brought into contact with a probe-immobilized substrate such as a probe-immobilized chip, and a hybridization reaction between the probe on the probe-immobilized substrate and the sample nucleic acid is performed. May be detected.

The present invention mainly comprises (1) a method using a labeling substance and (2) a means for detecting a hybridization reaction between the probe on the probe-immobilized chip and a nucleic acid component in a sample. Two types of electrochemical methods may be included.

In the case of the method (1) using a labeling substance, the sample nucleic acid is a fluorescent dye such as FITC, Cy3, Cy5 or rhodamine, or biotin, hapten,
It is labeled with an enzyme such as oxidase or phosphatase, or an electrochemically active substance such as ferrocene or quinones. Alternatively, detection is performed by using a second probe labeled with the aforementioned substance. A plurality of labeling substances may be used simultaneously.

In the case of the electrochemical method (2), a conductive substance is used as a substrate for immobilizing a probe, and a probe-immobilized tip is used as an electrode. Detection of the presence of a hybridization reaction using this electrode uses a counter electrode and a reference electrode in addition to this electrode, similarly to other general electrochemical detection methods. When arranging the reference pole, for example,
A common reference electrode such as a silver / silver chloride electrode or a mercury / mercury chloride electrode can be used. Probes having different base sequences may be immobilized on different conductive substrates, respectively, to form a chip disposed on the same substrate or the like. Thereby, highly accurate measurement can be performed. In that case, it is detected which probe the electrochemical signal obtained from each electrode corresponds to.

In some embodiments, the hybridization reaction between a nucleic acid component extracted from a sample substance and a probe immobilized on a probe-immobilized chip is performed, for example,
Perform as follows. That is, the hybridization reaction solution has an ionic strength of 0.01 to 5 and a pH of 5-1.
Perform in buffer range 0. In this solution, dextran sulfate which is a hybridization promoter, and
Salmon sperm DNA, bovine thymus DNA, EDTA and a surfactant may be added. The extracted nucleic acid component is added thereto and heat denatured at 90 ° C. or higher. Insertion of the probe-immobilized chip may be performed immediately after denaturation or after rapid cooling to 0 ° C. Further, it is also possible to carry out a hybridization reaction by dropping a solution on a substrate. During the reaction,
The reaction rate may be increased by an operation such as stirring or shaking. The reaction temperature is, for example, in the range of 10 ° C to 90 ° C,
The reaction time may be 1 minute or more and about 1 night. After the hybridization reaction, the electrode is taken out and washed. For washing, for example, in the range of ionic strength 0.01-5, pH
Use a buffer in the range of 5-10.

In the case of the method (1) using a labeling substance, the detection of the hybridization reaction is carried out by using an appropriate detection device according to the kind of the label, using the labeled base sequence in the sample or the secondary probe in the secondary probe. This is done by detecting the label. When the label is a fluorescent substance, the label may be detected using, for example, a fluorescence detector.

In the case of the electrochemical method (2), detection is performed in the following procedure. After washing the substrate, a double-stranded recognizer that selectively binds to the double-stranded portion formed on the electrode surface is acted on to perform electrochemical measurement. Although the double-stranded recognizer used here is not particularly limited, for example, Hoechst 33258, acridine orange, quinacrine,
It is possible to use bisintercalators such as dounomycin, metallointercalators, and bisacridines, tris intercalators, polyintercalators, and the like. Furthermore, these intercalators can be modified with an electrochemically active metal complex such as ferrocene, viologen and the like. The concentration of the DNA binding substance varies depending on its type, but is generally used in the range of 1 ng / mL to 1 mg / mL. In this case, the ionic strength is in the range of 0.001 to 5 and the pH is 5 to 1
A buffer in the range of 0 is used. After the electrode is reacted with the double-stranded recognizer, the electrode is washed and electrochemically measured.

In the electrochemical measurement, a potential higher than the potential at which the double-stranded recognizer reacts electrochemically is applied, and the reaction current value derived from the double-stranded recognizer is measured. At this time, the potential can be swept at a constant speed, applied as a pulse, or a constant potential can be applied. In the measurement, the current and the voltage are controlled using a device such as a potentiostat, a digital multimeter, and a function generator. Based on the obtained current value, the concentration of the target nucleic acid can be calculated from the calibration curve.

The base sequence detecting apparatus using electrodes includes, for example, a nucleic acid extracting section, a nucleic acid reacting section, a double-stranded recognizer reacting section, an electrochemical measuring section, and a washing section.

Another example of a DNA chip based on an electrochemical technique is disclosed in, for example, the following document (Hashimoto et al. 1994, W.
ang et al. 1998). Hashimoto et al., DNA
We reported sequence-specific gene detection using a probe-modified gold electrode and an electrochemically active dye. The anodic current from the dye correlates with the concentration of the target DNA. Wang et al. Reported indicator-free electrochemical DNA hybridization. The configuration of this biosensor involves immobilization of an inosine-substituted probe (without guanine) on a carbon paste electrode and chronopotentiometric detection of duplex formation due to the presence of the guanine oxidation peak of the label. These documents are incorporated herein by reference.

The electrochemical technique does not require labeling of the sample nucleic acid. The detection is performed by measuring an electric signal. Therefore, complicated systems such as those required for fluorescence detection are not required. Therefore, such a method can also be expected to reduce the size of the system.

According to the embodiments of the present invention described above, a nucleic acid sequence relating to the above-mentioned disease possessed by the patient itself and a nucleic acid sequence of a pathogenic microorganism infected by the patient are obtained from a sample substance such as blood collected from an individual such as a patient. Information can be easily obtained.

The probe-immobilized substrate according to the embodiment of the present invention is used for predicting an appropriate treatment method in an individual exposed to a disease. The probe-immobilized substrate includes a second probe and a first probe immobilized on the same substrate. The second probe is a probe (second probe) for obtaining information on a nucleic acid sequence linked (linked) to the disease present in a chromosome in cells of an individual, and the first probe is a probe for the disease. (First probe) for obtaining information related to the nucleic acid sequence of the pathogenic microorganism that induces the phenotype. By virtue of the advantages of the probe-immobilized substrate, information related to the prediction of the responsiveness of the treatment of a specific disease and / or a specific disease derived from the individual and the pathogenic microorganism can be easily and easily obtained from the sample substance extracted from the individual. It is possible to get.

In some embodiments, the method and the probe-immobilized substrate can predict the therapeutic effect of hepatitis C. Another aspect evaluates AIDS treatment (eg, HIV quantification and mutation detection, etc.)
And the probe-immobilized substrate used for the method. A further aspect is the method and probe-immobilized substrate used to evaluate treatment of hepatitis B (eg, quantification of HBV, detection of type and mutation, etc.).

For example, it is known that infection with hepatitis C progresses to liver cancer via cirrhosis. One of the treatment methods is IFN treatment in which interferon (hereinafter, referred to as IFN) is administered. Many use IFNα and β for therapy. However, it is known that the effectiveness of IFN varies widely. For example, IFN treatment is effective only in about 20 to 30% of Japanese human patients, and even if effective, very strong side effects have been reported. The difference in the therapeutic effect due to such individual differences is due to the hepatitis C virus (hereinafter referred to as HCV) infected by the individual.
Genotype (J. Clin. Microbiol.,
34, 2516 (1996)) and viral load;
It is thought to be due to individual differences in the nucleic acid sequence of the individual.

At least six major genotypes have been reported based on a comparison of the nucleotide and predicted amino acid sequences of different regions of HCV (Forns et al., J. Cl.
in.Microbiol., 34, 2516 (1996), Andonov et al.,
J. Clin. Microbiol., 33, 254 (1995).

Thus, according to aspects of the present invention, it is possible to predict the effect of IFN treatment on hepatitis C. Here, in the present specification, “interferon (I
“FN)” is used as a term to indicate interferon α, β, γ and / or ω. For example, IFN treatment is less effective when the infected virus is type 1b (common in Japanese). This is due to HCV-RNA in serum,
It means that HCV antibodies, GOP and GPT are not reduced by IFN therapy. In the case of type 2a, the effect of the treatment is higher. This is due to the serum HCV-RNA, H
Mean that CV antibodies, GOP and GPT are reduced by IFN therapy. In addition, when the viral load is 10 ⁶ cop
The effect is small even when it is as large as y / mL or more. Therefore, a first probe is used to determine the genotype and viral load of the infected virus at the nucleic acid level.

For example, as a probe used for detecting the presence of a nucleic acid sequence of hepatitis C virus (hereinafter also referred to as HCV) in a sample substance, a base corresponding to HCV-RNA, a fragment thereof, or a complementary sequence thereof is used. Sequences. A probe according to an embodiment of the present invention desirably comprises H
At least 11 bases, preferably at least 12 bases, more preferably at least 13, 14, 15, 16, 1 for detecting genotype or mutation of CV-RNA.
7, 18, 19, 20, 21, 22, 23, 24, 2
It has a sequence of 5, 26, 27, 28, 29 and 30 bases. Alternatively, the probe has at least 3
0, preferably about 40, 50, 60, 70, 80, 9
Even longer than 0, 100, 200, 500, 1000 bases are preferred for HCV genome detection. 11 to 30
Probe lengths longer than 30 bases are suitable for general hybridization formation. For example, SEQ ID NOs: 1 and 2 shown in the sequence listing
Those having a base sequence corresponding to 3, 4, or a complementary sequence thereof can be used. These arrays are
Most conserved among all HCV genotypes (Andonov et al., J. Clin. Microbiol., 33, 254 (1
995).

It is also preferable to use a probe for detecting not only the presence of the pathogenic microorganism in the sample substance but also a gene mutation of the pathogenic microorganism.
In the case of hepatitis C, it has been reported that the effects of IFN treatment differ depending on the mutation at a specific site in the nucleic acid sequence of the virus (Nagayama et al., Hepatology, 31, 745 (200
0)). An example of a first probe for detecting a mutation at a specific site in a nucleic acid sequence of a virus in a sample substance is Hepat
HCV polyproteins 434, 580, described in J.Eg., 31, 745 (2000);
It is possible to use a probe having a nucleotide sequence that determines the difference between the amino acids at positions 938, 962, 1176, and 2774. Hepatocellular carcinoma (HCC)
The genomic sequence of HCV-1b from patients tends to have more mutated residues than from asymptomatic carriers (hereinafter ASC). By using these, highly accurate prediction becomes possible.

As described above, depending on the genotype of HCV, I
Different effects of FN treatment have been reported. Therefore, the first probe may be a base sequence corresponding to HCV-RNA, a fragment thereof, or a sequence complementary thereto. For example, a probe that specifically detects a genotype of HCV type 1 (SEQ ID NO: 5), type 2 (SEQ ID NO: 6), and type 3 (SEQ ID NO: 7) is used as a first probe according to the genotype to be detected. May be used. That is, a nucleic acid having a sequence corresponding to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or their complementary sequence shown in the sequence listing,
It may be used as a probe.

When a probe capable of detecting a genotype is used, highly accurate detection is possible by simultaneously immobilizing a plurality of probes corresponding to different genotypes on a substrate.

In order to detect the genotype of the pathogenic microorganism in the sample substance, in one embodiment, PCR is performed on the nucleic acid sequence of the pathogenic microorganism in the sample substance using primers characteristic of the genotype in advance. It is also possible to carry out the reaction and then detect it with an immobilized probe, a universal probe capable of detecting all hepatitis C viruses without regard to their genotype (Andonov et al.
al., J. Clin. Microbiol., 33, 254 (1995).

In the case of hepatitis C, a sense strand having the nucleotide sequence shown in SEQ ID NO: 8 or 9 is used, and as an antisense strand, SEQ ID NO: 1 is used for type 1a.
SEQ ID NO: 11 for types 0 and 1b, SEQ ID NO: 1 for types 2a
SEQ ID NO: 13 for type 2, 2b, SEQ ID NO: 14 for type 3a
A PCR reaction may be performed using the nucleotide sequence of SEQ ID NO: 15, and the universal probe having the nucleotide sequence of SEQ ID NO: 15 may be used.

The amount of the pathogenic microorganism in the sample substance is also measured using the above-mentioned first probe, that is, a probe having a nucleotide sequence corresponding to a nucleic acid sequence derived from the pathogenic microorganism, a fragment thereof, or a complementary sequence thereof. It is possible.

For example, in the case of hepatitis C, it has been reported that the effect of IFN therapy differs depending on the amount of HCV in the blood (J. Clin. Microbiol., 33,
254 (1995)). INF, if HCV that patient has a serum exceeds 10 ⁶ copies / mL, obtained an effect difficult (Yeh et al., J. Med . Biol., 66, 481
(2002). Therefore, a probe for quantitatively evaluating the amount of virus includes a nucleotide sequence corresponding to HCV-RNA, a fragment thereof, or a complementary sequence thereof. When the nucleic acid in the sample substance is fluorescently labeled, the concentration of the virus is calculated from the fluorescence intensity of the label on the substrate after the probe on the chip and the nucleic acid in the sample substance are hybridized. When the electrochemical method is used, after the probe on the chip is hybridized with the nucleic acid in the sample substance, the concentration is calculated from the current value obtained from the chip.

In the probe-immobilized chip of the present invention,
The probe described above as the first probe, SEQ ID NOS: 5, 6
And / or 7 can be used to obtain information such as the genotype or amount of the virus of the pathogenic microorganism infected by the individual, the mutation at a specific site, the amount and quality of the expressed gene, and the like. If the viral load is as high as 10 ⁶ copy / mL, the effect of IFN therapy is small, but if the viral load is as high as 10 ⁶ copy / mL, the effect of IFN therapy is small.
The effect of the FN therapy can be predicted to be low, thereby predicting the effectiveness of the IFN therapy on the individual.

Another method for predicting the effect of IFN treatment on hepatitis C is disclosed in Japanese Patent Application No. 2001-62371.
And Japanese Patent Application No. 2001-62372. That is, a specific single nucleotide polymorphism (single nucleotide) present at the -88 position (hereinafter referred to as MxA-88) of the promoter region of the gene encoding the MxA protein of the human gene, that is, the MxA promoter region.
polymorphism (hereinafter referred to as SNP) is I
This is a method of predicting the magnitude of the IFN treatment effect by examining the SNP in order to determine the magnitude of the FN treatment effect. That is, INF is a SNP in the MxA promoter region.
Is effective in patients having a particular type of Therefore, by checking for the presence of the SNP,
The effect of FN therapy is evaluated. Therefore, in one embodiment of the present invention, a second probe for detecting an SNP in MxA is used as the second probe for detecting an individual nucleic acid.

According to the above findings, the position of -88 of the MxA promoter region (hereinafter referred to as MxA-88) is G / A.
In the case of G type, IFN therapeutic effect is low, and G / T and T
/ T type has high therapeutic effect, -12
Based on the fact that the 3rd position (hereinafter referred to as MxA-123) is of C / C type, the therapeutic effect of IFN is low, and that of C / A and A / A type, the therapeutic effect of IFN is high. Predict. Here, the notations “−88” and “−123” are positions when the transcription start site of the MxA gene is set to +1. FIG. 1 of WO 01/71001 shows the nucleotide sequence of the promoter region of the MxA gene.

Therefore, when predicting the therapeutic effect of IFN on hepatitis C according to the embodiment of the present invention, for example, H
A first probe for detecting the presence of the CV gene, furthermore, a first probe for detecting the genotype or mutation of HCV, and a second probe for detecting the type of base in the SNP of the promoter region of the gene encoding the MxA protein of the individual. By using a probe-immobilized chip immobilized with a probe, by performing a genetic diagnosis of HCV in a sample substance collected from an individual and a genetic diagnosis of a promoter region of a gene encoding an MxA protein possessed by the individual,
The effect of the IFN treatment can be easily predicted.

Further, as a method for predicting the effect of IFN treatment on hepatitis C, the magnitude of the IFN treatment effect is predicted based on the two SNP types of the mannose-binding lectin (hereinafter referred to as MBL) gene. Methods are known (M. Matsushita et a
l. , J. et al. Hepatology, 29; 695-
700, 1998). MBL is an important factor of the innate immune system, and genetic polymorphism exists. Group “XB” of the MBL genotype does not respond to IFN. Rosen reports on the sequence of MBL (Immunology, 9
3, 421 (1998)). This document is incorporated herein by reference. Therefore, genetic diagnosis of the mannose-binding lectin (hereinafter, referred to as MBL) gene may be performed together with or instead of performing the genetic diagnosis of the promoter region of the gene encoding the MxA protein as described above.

The polymorphic sites of MBL that affect the effect of IFN are located at position −221 of the promoter region of the MBL gene (hereinafter referred to as MBL-221) and at codons 52, 54 and 57 of exon 1 of the MBL gene. Exists. Here, the notation of “−221 position” is a position when the transcription start site of the MBL gene is set to +1.

The possible bases of the MBL-221 genotype are C or G. In the case of C, the base is called type X,
In the case of G, it is referred to as type Y (this nomenclature follows Madsen et al .; Madsen HO, Garred P,
Thiel S, Kurtzhals JAL,
See Lamm LU, Ryder LP, et al.
"Interaction between promoter and structural gene controls the lowest serum level of human mannan binding protein." J Immunol 1995; 155: 3013.
-20). This document is incorporated herein by reference.

Codons 52, 54 and 57 of exon 1 of the MBL gene are present in the collagen-like domain which is a structural gene. The possible genotype of codon 52 is CGT or TGT. According to the classification of Madsen et al., The former is type A and the latter is type D. Similarly, the possible genotype of codon 54 is GGC or GGC.
According to the classification, the former is type A, and the latter is type B. The genotype that codon 57 can take is GGA or GAA. According to the same classification, the former is type A and the latter is type C. In all these codons, type A is the wild type allele and the other types B, C and D are mutant alleles. Codon 5
If the 2, 54 and 57 alleles are all type A, the effect of IFN is higher than in the other case, ie, when at least one of the alleles is not type A.

The MBL-221 is of type Y and the alleles of codons 52, 54 and 57 are of type A
In the case of YA-YA homozygous patients, the effect of IFN is more likely to be obtained than in other types of patients. The MBL-221 is of type Y, and any of the alleles of codons 52, 54 and 57 is of type A.
In other words, in the case of a patient having YnonA-YA heterozygous or YnonA-YnonA homozygous, the effect of IFN is less likely to be obtained than in the case of YA-YA homozygous patient.

From the above findings, it is possible to determine the second probe.

The disease is hepatitis C, and the drug is I
In one embodiment that is FN, the second probe used in hybridization to the nucleic acid of the individual (including use on a probe-immobilized chip) is a promoter that detects the SNP of the MxA promoter as described above. If the disease is hepatitis C and the drug is IF
In other embodiments that are N, the second probe used in hybridization to the nucleic acid of the individual (including use on a probe-immobilized chip), as described above, has a MBL that is associated with the effect of IFN. Probe for detecting polymorphism.

Each of these nucleotide sequences in the promoter region of the human MxA gene, including the SNPs at positions 455 and 420, is involved in the effects of IFN therapy. Accordingly, in a further embodiment of the base arrangement set forth in SEQ ID NOs: 16, 17, 18, 19, 37, 38, 39 and 40,
The probe is selected from the group consisting of: (at455) a nucleotide sequence represented by SEQ ID NO: 16 in the following sequence listing; (bt455) several nucleotides excluding the nucleotide at position 455 in the nucleotide sequence represented by (at455); A modified base sequence in which bases are deleted, substituted, or added with one or more bases; ability of a modified probe for detecting MBL polymorphism to hybridize to an individual nucleic acid encoding MBL Has been maintained.
The hybridization conditions used for the modified probes are different from those of general probes. Desirably, the modified probes according to the present invention are HCV-
At least 11 bases, preferably at least 12 bases, more preferably at least 13, 14, 15, 16, 17, 1 for detecting RNA genotype or mutation.
8, 19, 20, 21, 22, 23, 24, 25, 2,
It has a sequence of 6, 27, 28, 29 and 30 bases.
Alternatively, the probe is at least 30, preferably about 40, 50, 60, 70, 80, 90, 10,
Even if longer than 0, 200, 500, 1000 bases, H
Preferred for detection of CV genome. Lengths from 11 to 30 are suitable for chip formation, and probes longer than 30 bases are suitable for general hybridization formation.

(Ct455) a base sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 16, (dt455) a base sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 16, (et455) ~ (Dt455) the complementary sequence of the base sequence selected from (ag455) the base sequence shown in SEQ ID NO: 17 of the following sequence listing, (bg455) excluding the base at position 455 in the base sequence shown in (ag455) A modified base sequence in which several bases have been deleted, substituted or added with one or more bases, (cg455) a base sequence containing the sequence shown in positions 441 to 455 of SEQ ID NO: 17, (dg455) SEQ ID NO: 17 A nucleotide sequence comprising a sequence shown at positions 449 to 459 of (eg455) a complementary sequence of a nucleotide sequence selected from the (ag455) to (dg455) (aa455) a nucleotide sequence represented by SEQ ID NO: 18 in the following sequence table, (ba455) Several bases except for the base at position 455 in the base sequence represented by the above (aa455) have been deleted or substituted, or one or more bases have been added. A decorative nucleotide sequence, (ca455) a nucleotide sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 18, (da455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 18, (ea455) the (aa455) ) To (da455), a complementary sequence of a base sequence selected from (ac455) a base sequence represented by SEQ ID NO: 19 in the following sequence listing, (bc455) excluding a base at position 455 in the base sequence represented by (ac455) A modified base sequence in which several bases have been deleted, substituted or added with one or more bases, (cc455) a base sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 19, (dc455) SEQ ID NO: 19 A base sequence selected from the group consisting of: a base sequence containing the sequence shown at positions 449 to 459 of (ec455), a complementary sequence of the base sequence selected from (ac455) to (dc455). .

SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 1
8, SEQ ID NO: 19, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40 are nucleotide sequences including the promoter region of the human MxA gene, and positions 455 and 420 of these nucleotide sequences. S exists in
The nucleotide sequences of SEQ ID NOS: 16 to 19, in which NP is involved in responsiveness to the effects of IFN therapy, have a common sequence except for the nucleotide at position 455. Position 455 of SEQ ID NO: 16 is thymine. Position 455 of SEQ ID NO: 17 is guanine. It is adenine at position 455 of SEQ ID NO: 18. Position 455 of SEQ ID NO: 19 is cytosine.

An HCV-infected person having the nucleic acid sequence of SEQ ID NO: 16 in which these bases at position 455 are thymine is IFN-infected.
HCV-infected individuals who do not have the nucleic acid sequence of SEQ ID NO: 16, where the base at position 455 is thymine, are not effective with IFN therapy. That is, HCV having a heterozygous nucleic acid of SEQ ID NO: 16 in which the nucleotide sequence at position 455 is thymine and a nucleic acid of SEQ ID NO: 17 in which the nucleotide sequence at position 455 is guanine (hereinafter abbreviated as G / T heterozygous).
The nucleotide sequence at position 455 is guanine as compared to an infected person or an HCV-infected person having the nucleic acid of SEQ ID NO: 16 in which the nucleotide sequence at position 455 is thymine homozygously (hereinafter abbreviated as T / T homologous). HCV-infected individuals having the nucleic acid of SEQ ID NO: 17 homozygously (hereinafter abbreviated as G / G homologous)
N therapy is not effective.

Alternatively, T / non-T hetero or T /
Compared to T homozygous HCV infected persons, HCV infected persons having a homozygous promoter region of the MxA gene whose nucleotide sequence at position 455 is not thymine (hereinafter referred to as non-T / non-
(Abbreviated as T-homo) showed that IFN therapy was not effective. Examples of non-T / non-T homo combinations include G / G, G / A, G / C, A / A, A / C, and C / G.
There is C. As a combination of T / non-T, T /
G, T / A, and T / C.

The nucleotide sequences of SEQ ID NOs: 37 to 40 have a common sequence except for the nucleotide at position 425. 42 of SEQ ID NO: 37
Position 5 is adenine. Position 425 of SEQ ID NO: 38 is cytosine. In addition, position 425 of SEQ ID NO: 39 is thymine. Position 425 of SEQ ID NO: 40 is guanine.

In the probe-immobilized chip of the present invention,
As a second probe for detecting the sequence of a nucleic acid possessed by an individual, the nucleotide sequence of SEQ ID NO: 16 including the SNP site and having a nucleotide sequence of thymine, a fragment thereof, or a complementary sequence thereof ((at455) to (et455)) ) Can be used to determine whether or not the SNP site of the nucleotide sequence including the promoter region of the human MxA gene possessed by an individual is thymine before treatment. Thereby, the effectiveness of IFN therapy for the individual can be predicted.

Therefore, prior to the implementation of IFN therapy, for example, by determining the base of the SNP site in the base sequence including the promoter region of the human MxA gene possessed by an HCV infected patient, IFN therapy can detect efficacy.

Further, in the probe-immobilized chip of the present invention, as a second probe for detecting a sequence of a nucleic acid possessed by an individual, the nucleotide sequence of SEQ ID NO: 17 containing the SNP site and the nucleotide sequence of the SNP site is guanine , Its fragments or their complementary sequences ((ag455)-(eg4
55)) or the nucleotide sequence of SEQ ID NO: 18 wherein the nucleotide sequence of the SNP site is adenine, a fragment thereof or a complementary sequence thereof ((aa455) to (ea455)), or a sequence wherein the nucleotide sequence of the SNP site is cytosine No. 19, its fragment or its complementary sequence ((ac4
55) to (ec455)), it is possible to examine the sequence of the SNP site in the nucleotide sequence including the promoter region of the human MxA gene possessed by an individual, thereby predicting the effectiveness of IFN therapy for the individual. be able to. (Japanese Patent Application Nos. 2001-62371 and 20)
01-62372).

The disease is hepatitis C, and the drug is I
In one embodiment which is FN, the second probe used in hybridization to the nucleic acid of an individual (including use on a probe-immobilized chip) is as follows: (aa420) SEQ ID NO: 37 in the following sequence listing. Base sequence, (ba420) a modified base sequence in which several bases except the base at position 420 in the base sequence shown in (aa420) are deleted, substituted, or added with one or more bases, (ca420) (Da420) a complementary sequence of a base sequence selected from (aa420) to (ca420) (ac420) represented by SEQ ID NO: 38 in the following sequence table Base sequence, (bc420) a modified base sequence in which several bases except the base at position 420 in the base sequence shown in (ac420) are deleted, substituted, or added with one or more bases, (cc420) A base sequence comprising the sequence shown at positions 415 to 425 of SEQ ID NO: 38, (dc420) the (ac420) to ( cc420) the complementary sequence of the base sequence selected from (at420) the base sequence shown in SEQ ID NO: 39 of the following sequence listing, (bt420) several nucleotides excluding the base at position 420 in the base sequence shown in (at420) Bases deleted, substituted, or modified base sequence to which one or more bases have been added, (ct420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 39, (dt420) the above (at420) to (at420) (ct420) the complementary sequence of the base sequence selected from (ag420) the base sequence shown in SEQ ID NO: 40 of the following sequence listing, (bg420) several nucleotides excluding the base at position 420 in the base sequence shown in (at420) Bases are deleted, substituted, or modified base sequence to which one or more bases are added, (cg420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 40, (dg420) the (ag420) to ( cg420), a base sequence selected from the group consisting of the complementary sequence of the base sequence selected from cg420) may be used.

An HCV-infected person having the nucleic acid sequence of SEQ ID NO: 37 in which the base at position 425 is adenine can
HCV-infected individuals who do not have the nucleic acid sequence of SEQ ID NO: 37, in which the base at position 425 is adenine, are effective against N therapy, whereas IFN therapy is not effective. Details regarding sequence and effectiveness may be understood as described above for the SNP at position 455.

Further, when the disease is a disease for which IFN therapy has been confirmed to be effective, for example, hepatitis C, the second probe may be, for example, a sequence as described in (i) to (t) below. It may be. These are MBL-221
Are suitable nucleic acids for use in determining the genotype of and the presence of polymorphisms at codons 52, 54 and 57.

The MBL gene containing the polymorphisms encoded by MBL-221 and codons 52, 54 and 57 are shown in SEQ ID NOs: 41 to 56. MBL-221 is at position 425 of these sequences.

Exon 1 also contains 646 of these sequences.
Codon 52 changes from 868 to 870,
Codon 54 is at positions 874 to 876, and codon 57 is at positions 883 to 885. The SNP at codon 52 is at position 868, the SNP at codon 54 is at position 875, and the SNP at codon 57 is at position 884.

(I) Nucleic acid fragments of SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and SEQ ID NO: 44 containing the aforementioned SNP site of MBL-221, wherein SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and SEQ ID NO: Number 44, 418-432
A fragment containing a nucleic acid having a sequence corresponding to a position.

(J) Nucleic acid fragments of SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and SEQ ID NO: 44 containing the SNP site of MBL-221, wherein SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, Number 44 421-430
A fragment containing a nucleic acid having a sequence corresponding to a position.

(K) The complementary sequence according to the above (i) and (j).

(L) Codons 52, 54 and 57
SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 5
0, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, including nucleic acids having sequences corresponding to positions 868 to 885 of these sequences. fragment. For example, SEQ ID NO: 4
A fragment comprising the nucleic acid set forth in SEQ ID NO: 5 to SEQ ID NO: 56.

(M) SN of codons 52 and 54
SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 4 including P site
7, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 5
4. Fragments of the nucleic acids of SEQ ID NO: 55 and SEQ ID NO: 56, comprising a nucleic acid having a sequence corresponding to positions 868 to 876 of these sequences. For example, a fragment containing the nucleic acid shown in SEQ ID NO: 45 to SEQ ID NO: 56.

(N) SN of codons 54 and 57
SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 4 including P site
7, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 5
4. Fragments of the nucleic acids of SEQ ID NO: 55 and SEQ ID NO: 56, comprising a nucleic acid having a sequence corresponding to positions 874 to 885 of these sequences. For example, a fragment containing the nucleic acid shown in SEQ ID NO: 45 to SEQ ID NO: 56.

(O) SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 including the SNP site of codon 54
A fragment of the nucleic acid of SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, or SEQ ID NO: 56, comprising a nucleic acid having a sequence corresponding to positions 874 to 876 of these sequences. For example, SEQ ID NO: 45 to SEQ ID NO: 56
A fragment containing the nucleic acid shown in (1). In particular, a fragment containing positions 869 to 880 is desirable.

(P) SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 including the SNP site of codon 52
A fragment of a nucleic acid of SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO: 56, comprising a nucleic acid having a sequence corresponding to positions 868 to 870 of these sequences. For example, SEQ ID NO: 45 to SEQ ID NO: 56
A fragment containing the nucleic acid shown in (1). In particular, a fragment containing positions 864 to 873 is desirable.

(Q) SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 including the SNP site of the codon 57
A fragment of the nucleic acid of SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO: 56, comprising a nucleic acid having a sequence corresponding to positions 883 to 885 of these sequences. For example, SEQ ID NO: 45 to SEQ ID NO: 56
A fragment containing the nucleic acid shown in (1). In particular, a fragment containing positions 880 to 890 is desirable.

(R) SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 including the SNP site of the codon 54
A fragment of the nucleic acid of SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, wherein the fragment comprises the nucleic acid containing position 875 of the sequence where the SNP of the codon 54 is present. For example, SEQ ID NO: 45 to SEQ ID NO: 5
A fragment containing the nucleic acid shown in 6.

(S) A nucleic acid fragment shown in (i) to (m), wherein the length is 11 to 30. Preferred probe sequences are at least positions 420 to 430, 864 to 874, 870 to 880 and 87 for the MBL gene.
Includes positions 9 through 889. As for the MxA gene, it is preferable to include at least positions 450 to 460.

(T) A nucleic acid fragment shown in (i) to (m) above, wherein one or several nucleotides excluding the polymorphic site are deleted, substituted, or added.

In each sequence described in the sequence listing of the present specification,
“N” and “n” indicate any base of adenine, thymine, guanine or cytosine.

In the probe-immobilized chip of the present invention,
The measurement of nucleic acid (RNA) expressed by an individual can also be performed using the above-mentioned second probe, that is, a probe having a base sequence corresponding to a nucleic acid sequence derived from an individual, a fragment thereof, or a complementary sequence thereof. In this case, the nucleic acid chain is R
NA or cDNA or cRNA, which can be detected or quantified. For example, the effect of IFN differs depending on the constitution of an individual, and therefore, by measuring the gene expression pattern (quantity, quality) upon administration, it is possible to predict the efficacy.

As described above, the target disease according to the embodiment of the present invention is a disease induced by a pathogenic microorganism.
There is no particular limitation. The present invention includes diseases relating to pathogenic microorganisms. The invention also includes oncological diseases. For example, when predicting the effectiveness of IFN treatment,
Even if the disease other than hepatitis C is a disease for which the effectiveness of IFN treatment has been confirmed, the therapeutic effect can be similarly predicted. Such examples include the hepatitis virus (A, B, C, D,
E, F, G), HIV, influenza virus, herpes group virus, adenovirus, human polyoma virus, human papilloma virus, human parvovirus, mumps virus, human rotavirus, enterovirus, Japanese encephalitis virus, dengue virus, rubella virus And viral infections such as HTLV, Staphylococcus aureus, Streptococcus haemolyticus, pathogenic Escherichia coli, Vibrio parahaemolyticus,
Helicobacter pylori, Campylobacter, Cholera, Shigella, Salmonella, Yersinia, Neisseria gonorrhoeae, Listeria, Leptospira, Legionella, Spirochetes,
Examples include, but are not limited to, bacterial infections such as mycoplasma pneumonia, rickettsia, chlamydia, malaria, dysentery amoeba, and pathogenic fungi, parasites, and fungal diseases.

Furthermore, hereditary diseases, retinoblastoma, Wilms tumor, familial polyposis of the large intestine, hereditary non-polyposis colorectal cancer, neurofibromatosis, familial breast cancer, xeroderma pigmentosum, brain tumor, oral cancer , Esophageal cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, lung cancer, thyroid tumor, breast tumor, urinary tumor, male organ tumor, female organ tumor, skin tumor, bone / soft tissue tumor, leukemia, lymphoma and solid tumor It is also possible to predict the efficacy of IFN treatment for such neoplastic diseases.

The pathogenic microorganism that can be detected according to the embodiment of the present invention is not particularly limited. For example, when predicting the efficacy of IFN treatment, it induces a disease for which the effectiveness of IFN treatment has been confirmed. Pathogenic microorganisms include, for example, HCV, but are not particularly limited as long as they are pathogenic microorganisms related to diseases for which the effectiveness of IFN treatment has been confirmed. If the pathogenic microorganism is directly or indirectly correlated with the disease or symptoms of the disease, the pathogenic microorganism is associated with the disease.

For example, hepatitis viruses (A, B, C, D,
E, F, G), HIV, influenza virus, herpes group virus, adenovirus, human polyoma virus, human papilloma virus, human parvovirus, mumps virus, human rotavirus, enterovirus, Japanese encephalitis virus, dengue virus, rubella virus And viruses such as HTLV, Staphylococcus aureus, hemolytic streptococci, pathogenic Escherichia coli, Vibrio parahaemolyticus, Helicobacter pylori, Campylobacter, cholera, Shigella, Salmonella, Yersinia, Neisseria gonorrhoeae, Listeria,
It can be used for the detection of bacteria, parasites and fungi such as Leptospira, Legionella, Spirochetes, Mycoplasma pneumonia, Rickettsia, Chlamydia, Malaria, Shigella amoeba and pathogenic fungi.

In the above example, an example of IFN therapy and analysis of each disease and individual gene has been described. However, the present invention is not limited to this. In some embodiments, for example, human immunodeficiency virus (hereinafter referred to as HIV) present in an individual
And by analyzing the individual gene, AIDS (acqu
It is possible to predict the susceptibility of ired immunodeficiency syndrome (AIDS). Furthermore, after analyzing the genotype and / or copy of HIV and a specific polymorphism of the gene of the individual, based on the obtained information, the following drugs, for example, ritonavir (hereinafter referred to as RTV) and saquinavir ( The therapeutic effect of a protease inhibitor such as SQV) and a reverse transcription inhibitor such as azidothymidine (hereinafter AZT) and didanosine (hereinafter referred to as ddl) may be predicted. In some further aspects,
Varicella Zost virus present in individuals
er Virus; VZV) and the gene of the individual can be used to predict the susceptibility of varicella or shingles (zona). Further, the effectiveness of an antiviral drug such as acyclovir may be analyzed from the obtained information on the nucleic acid. Alternatively, by analyzing the genotype and copy number of an influenza virus (influenza virus) and the genotype of an individual, the susceptibility to influenza and the efficacy of an antiviral drug such as Tamifulu may be analyzed.

When the extracted nucleic acid component is HCV-RNA, HCV-RNA may be directly used as a sample nucleic acid, or cVDN from HCV-RNA using reverse transcriptase.
After preparing A, it may be used as a sample nucleic acid.

When using a probe capable of detecting a genotype, a plurality of probes corresponding to different genotypes can be immobilized on a substrate at the same time and used to achieve highly accurate detection.

The following examples are intended to illustrate but not limit the present invention.

EXAMPLES Example 1 describes a method for extracting nucleic acids from a human subject Extraction of Human Genome and Virus Genome from Patient Patients Approximately 3 mL of blood from HCV-infected patients was collected in a vacuum blood collection tube containing EDTA2K. After that, QIAam
pDNA Blood Midi Kit (QIAG
EN) was used to perform nucleic acid extraction. A reverse transcription reaction was performed on the HCV genomic RNA contained in the obtained extract. This reverse transcription reaction is performed on a part of the extract,
Hexa-deoxyribonucleide
mixture (TAKARA) and M-MLVReve
rs Transcriptase (LIFE TE
CHNOLOGIES) at 42 ° C. for 30 minutes.

2. Examination of extraction efficiency when extracting HCV genomic RNA using human genome extraction kit When extracting HCV genomic RNA, serum is usually used instead of whole blood. Furthermore, after removing impurities such as unnecessary proteins as much as possible, HCV genomic RNA is extracted using a guanidine buffer or the like. However, in the method according to the embodiment of the present invention, simultaneous extraction with human genomic DNA is performed. Therefore, H in the case where human genomic DNA was extracted using a kit for human genome extraction using whole blood
The extraction efficiency of the CV genome was examined.

As a general method, SepeGeneRV-R
Method for extracting HCV genome from 100 μL of serum using (Sanko Junyaku) and QIAamp DNA Blood
The method of extracting the HCV genome from whole blood using dMidi Kit (QIAGEN) was compared. As for the method of extracting from whole blood, see QIAamp DNAB
After extracting the HCV genome with the load Midi Kit, the results obtained by subjecting the extracted product to a reverse transcription reaction were compared between a case where the extracted product was purified by ethanol precipitation and a case where ethanol precipitation was not performed. .

To evaluate the extraction efficiency, 5 ′ of the HCV genome was used.
Competitive PC using UTR sequence
R (K. Chayama et al., J. Ga.
stroenterol. Hepatol. 8, S
40-44, 1993).

Table 1 shows the results.

[0109]

[Table 1]

As a result, compared with the conventional HCV RNA extraction method, QIAamp DNA Blood Midi was used.
It was found that the method using Kit reduced the extraction efficiency of HCV RNA by about 10 times. Therefore, QIAa
The extraction method using the mp DNA Blood Midi Kit may not be able to be measured when performed on a sample having an extremely low blood virus amount, or may be used in applications such as quantification of virus using a chip. It was considered difficult to use. However, conversely, in a sample in which the amount of virus in a conventional method occupies a concentration of 10 copy / ml or more in whole blood, theoretically, QIAamp DNA Blood
It was suggested that simultaneous extraction of DNA and RNA using dMidi Kit was possible. That is, QIAam
It was suggested that the virus could be qualitatively measured using the extract using the pDNA Blood Midi Kit.

The samples obtained by performing the reverse transcription reaction as described above were subjected to simultaneous amplification of the human genome and the HCV genome.

For the primers for amplification, MxAF01 and MxAR02 were used for the human genome. HC
For the V genome, nc2 (K. Chayama et al.)
al. , J. et al. Gastroenterol. He
Patol. 8, S40-44, 1993, n
t27-45) and primer33 (Okamotoe
t al. Jpn J. et al. Exp. Med. 6
0, 215-222, 1990). Amplification was performed in one tube. PCR was performed at 94 ° C for 3 hours.
Fifty cycles of 0 second, 55 ° C. for 30 seconds, and 72 ° C. for 1 minute were performed for 50 cycles, followed by treatment at 94 ° C. for 4 minutes and then at 72 ° C. for 7 minutes.

Sequence of Primer: MxAF01: 5'-ACACACCCGTTTCCACCCTGGAGAGGCCAG-3 'MxAR02: 5'-TGCGCAGTGCTGGAGTGCGGCCTCCGCTCT-3' NC2: 5'-CCT GTG AGG AAC TAC TGT C-3 '33: 5'-GGT GCA CGG TCG ACG AGA CC-3 '.

As a result, the product derived from the human genome (siz
e: 610 bp) and a product derived from the HCV genome (si
ze: 300 bp) were both amplified. Therefore, it became clear that the process from the extraction of the genome to the amplification of the specific region can be achieved by performing the same treatment on both human genomic DNA and HCV genomic RNA.

According to the embodiment of the present invention, it is possible to easily and quickly predict a treatment method for the disease from a sample substance extracted from an individual. Further, according to such an embodiment, since a plurality of pieces of information can be obtained simultaneously from one sample obtained from an individual, it is possible to prevent a mistake in the sample that may occur during the test. Further, even when handling a sample having a high risk, it is possible to narrow the range of contamination by such a sample as compared with the conventional case, and to minimize the risk of a contamination accident caused by the sample. It is possible.

Example 2 describes a DNA chip for evaluating the effect of IFN therapy. A DNA chip for predicting the effect of IFN therapy First, human chromosomal DNA and H were obtained from the blood of a patient (human).
CV-RNA was collected. Chromosomal DNA is SEQ ID NO: 2
0, a 135 bp fragment was amplified using the primers of SEQ ID NO: 21. For HCV-RNA, cDNA was prepared using reverse transcriptase.

The nucleic acid sample obtained by the above steps was labeled with FITC using a kit manufactured by Pharmacia.

FIG. 1 shows a schematic diagram of the DNA chip of this embodiment. D of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26 as a second probe on a substrate 6 made of a slide glass coated with polylysine
DNA probes of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 (the first probe and the second probe are indicated by 1 to 5 in FIG. 1) were spotted at 200 nL each as the NA probe and the first probe, and dried. Thereafter, the probe was immobilized on the substrate 6 by performing UV irradiation. SEQ ID NOs: 22 to 25
Is a fragment containing the SNP site at position 455 of the nucleotide sequences of SEQ ID NOS: 16 to 19, respectively. SEQ ID NO: 5
The nucleotide sequences of Nos. 7 to 7 are HCV virus type 1 and 2 respectively.
It is a probe that detects genotypes 3 and 3.

[0119] The nucleic acid sample labeled with FITC was
It was dissolved in a C-1 mmol / L EDTA solution. This was placed in a reaction chamber having a volume of 20 μL, and covered with a probe-immobilized slide glass. 50 ° C, 1
After performing the reaction for 2 hours, 0.2 × SSC-1 mmol
After washing twice with the / LEDTA solution, the fluorescence on the slide glass was measured.

As a result, significant fluorescence was observed only from the spot where the probe of SEQ ID NO: 22 was immobilized. MxA-88 in human-derived nucleic acid contained in this sample substance
Was considered to be T / T homozygous. Also,
The virus contained in this sample substance was determined to be type 2. Further, from the obtained fluorescence intensity, the virus concentration was 10
It was estimated to be ⁶ copy / mL or less. Therefore,
It was expected that IFN would work effectively in patients from whom this sample material was collected.

Example 3 describes a PNA chip for evaluating the effect of IFN therapy. A PNA chip for predicting the effect of IFN therapy First, human chromosomal DNA and H were obtained from the blood of a patient (human).
CV-RNA was collected. Next, a 135 bp fragment of the chromosomal DNA was amplified using the primers of SEQ ID NOs: 20 and 21. For HCV-RNA, cDNA was prepared using reverse transcriptase. Using the obtained cDNA as a template, SEQ ID NO: 8 (this is a sense), SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14
A PCR reaction was performed using the primers (these are antisense).

FIG. 2 is a schematic diagram of the PNA chip of this embodiment. SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31 as a second probe having a N-terminal modified cysteine on a probe-immobilized chip in which ten gold electrodes 13 are patterned on a glass substrate 12. SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 3
Each of 6 PNA probes 7 to 11 was spotted by 200 nL and left for 1 hour.

The nucleotide sequences of SEQ ID NOS: 27 to 30 are fragments containing the SNP site at position 455 of the nucleotide sequences of SEQ ID NOs: 16 to 19, respectively. The nucleotide sequences of SEQ ID NOs: 32 to 36 are HCa virus types 1a, 1b, 2a, and 2a, respectively.
It is a probe that detects genotypes of b-type and 3a-type.

Next, 2 × SSC-1 mmol
/ L EDTA solution. This was placed in a reaction chamber having a volume of 20 μL, and covered with a probe-immobilized tip. After performing the reaction at 50 ° C. for 1 hour, the reaction was performed at 0.
Washed twice with 2 × SSC-1 mmol / LEDTA solution. Next, 10 μmol / L Hoechst 3325 was added thereto.
Eight solutions were added dropwise. A counter electrode and a reference electrode are separately provided, and Hoechst 33258 when a voltage is applied between the gold electrode and the counter electrode.
The oxidation current from was measured.

As a result, a significant current change was observed from the gold electrode 13 on which the probes of SEQ ID NO: 27, SEQ ID NO: 28 and SEQ ID NO: 34 were immobilized, and the MxA-88 gene in the human-derived nucleic acid contained in this sample substance The type was considered to be a G / T heterotype. Further, the virus contained in this sample substance was determined to be type 2a. Therefore, it was predicted that IFN would work effectively in the patient from whom this sample material was collected.

Example 4 In accordance with the present invention, infected HCV in hepatitis C infected persons
Virus type, infected MxA-88, MxA-12
3. From the genotype of MBL-221 and the codon 54 of the collagen-like domain of MBL, it is possible to predict the efficacy of IFN therapy in hepatitis C infected individuals.

The type of virus, MBL, will be described with reference to FIG.
An example of a method for predicting the effect of IFN therapy from the genotype of MxA and MxA will be described.

In step 3a, the practitioner collects a sample such as a blood sample from an individual to whom IFN is to be administered, and starts prediction. The collected sample is subjected to processing such as purification and extraction as necessary, and then the type of virus and the genotype of MBL and MxA, that is,
MxA-88 and MxA-123 genotypes, MBL-2
Determine the genotype of 21 and the genotype of codon 54 of the collagen-like domain of MBL and proceed to step 3b.

In step 3b, if the type of virus determined in step 3a includes type 1, it is determined to proceed to step 3c, and if it is only type 2, it is determined to proceed to (3d).

In step 3c, the genotype and IxA are determined based on the MBL and MxA genotypes determined in step 3a.
The table 2 in which the effect of the FN is associated is searched to extract the effect of the corresponding IFN, thereby predicting the effectiveness of the IFN therapy, and terminating the entire prediction process.

At step 3d, based on the genotypes of MBL and MxA determined at step 3a,
A search is performed on table 3 in which the effects of FN are associated, and
Extract the effects of FN, thereby predicting the efficacy of IFN therapy, and end the entire prediction process.

The tables used here are genotypes and I
This is information associated with FN sensitivity. Each table is shown as a table, which is a table corresponding to each table number. Each table used in the method described above is provided as an example. The tables and tables used here may be those which associate each genotype with the effect of IFN. For example, Table 2 is a table showing the relationship between the genotype and the effect of IFN in patients infected with type 1 HCV. On the other hand, Table 3 is a table showing the relationship between the genotype and the effect of IFN in patients infected with type 2 HCV. Further, the table used may be a table containing only any one of the components, or may be a table composed of components in other combinations. In the above example, Tables 2 and 3 are used as tables referred to in Steps 3c and 3d, respectively, and two tables prepared by selecting necessary items in advance for each step are used. However, a single table prepared by combining them may be used. Alternatively, it may be a table composed of components of other combinations. For the selection of the components, for example, in the table used in step 3c, information indicating the association between the genotype and the significant or ineffective in the type 1 HCV infected person is selected,
In step 3d, information indicating the relationship between the genotype and the significant or ineffective response in HCV type 2 infected persons may be selected. However, the present invention is not limited to this, and the practitioner may arbitrarily select the components.

The numerical values of the components described in Tables 2 and 3 used here are an example of display indicating information in which the genotype is associated with IFN sensitivity or IFN effect. It is not limited to the numerical values described. That is, if it is a notation that can substantially show the correlation between genotype and significant or non-great effect, for example,
The score may be “○”, “×”, “△”, or may be a simplified numerical score (for example, 1, 2, 3, 4, and 5).

[0134]

[Table 2-1]

[0135]

[0136]

[Table 2-2]

[0137]

[Table 3]

According to the above-described embodiment of the present invention, a more accurate prediction can be made by examining many items.

By using the method and the probe-immobilized chip according to the above embodiment of the present invention,
The target nucleic acid information can be easily, quickly, and accurately.
It is possible to collect.

The above-described embodiment is an example of the present invention and is not described for the purpose of limiting the present invention.

Further benefits and modifications will be readily apparent to those skilled in the art. Accordingly, the invention in its broader aspects is not limited to the details and representative embodiments shown and described herein. Accordingly, various changes may be made without departing from the spirit or the general inventive concept as defined by the appended claims and their equivalents.

[0142]

According to the present invention, there has been provided a method capable of easily and accurately recovering target nucleic acid information from a sample substance in a short time.

[0143]

[Sequence List] SEQUENCE LISTING <110> KABUSHIKI KAISHA TOSHIBA <120> DETECTION OF NUCLEIC ACID ASSOCIATED WITH DISEASE <130> A000200297 <150> JP 2001-90053 <151> 2001-3-27 <160> 72 <210> 1 <211> 21 <212> DNA <213> Hepatitis C Virus <400> 1 ccctgtgagg aactwctgtc t 21 <210> 2 <211> 21 <212> DNA <213> Hepatitis C Virus <400> 2 ggtgcacggt ctacgagacc t 21 <210> 3 <211> 26 <212> DNA <213> Hepatitis C Virus <400> 3 tctagccatg gcgttagtry gagtgt 26 <210> 4 <211> 26 <212> DNA <213> Hepatitis C Virus <400> 4 cactcgcaag caccctatca ggcagt 26 <210> 5 <211> 18 <212> DNA <213> Hepatitis C Virus <400> 5 cgctcaatgc ctggagat 18 <210> 6 <211> 18 <212> DNA <213> Hepatitis C Virus <400> 6 cactctatgc ccggccat 18 <210> 7 <211> 18 <212> PNA <213> Hepatitis C Virus <400> 7 cgctcaatac ccagaaat 18 <210> 8 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 8 cgcgcgacta ggaagacttc 20 <210> 9 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 9 cgcgcgacgc gcaaaacttc 20 <210> 10 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 10 tgccttgggg ataggctgac 20 <210> 11 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 11 gagccatcct gcccacccca 20 <210> 12 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 12 gccccatgaa gggcgagaac 20 <210> 13 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 13 accctcgttt ccgtacagag 20 <210> 14 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 14 gctgagccca ggaccggtct 20 <210> 15 <211> 20 <212> DNA <213> Hepatitis C Virus <400> 15 aggaagactt ccgagcggtc 20 <210> 16 <211> 581 <212> DNA <213> Homo sapiens <400> 16 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgn 420 aggtgcgggg ccaggagcta ggtttcgttt ctgctcccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgc gac gggcg gacgggcg gacgggcg gagggggccag 540g <210> 17 <211> 581 <212> DNA <213> Homo sapiens <400> 17 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgn 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcgcccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcgggggag aaagaggcgc gacagggcg g aaagaggcgg aaagaggcgc ag <210> 18 <211> 581 <212> DNA <213> Homo sapiens <400> 18 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgn 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcacccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgac cc gg gag cc gg gg gg cc <210> 19 <211> 581 <212> DNA <213> Homo sapiens <400> 19 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgn 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcccccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgg gacgggcg gacgggcg gacgggccag cc <210> 20 <211> 22 <212> DNA <213> Homo sapiens <400> 20 aggtgcgggg ccaggagcta gg 22 <210> 21 <211> 22 <212> DNA <213> Homo sapiens <400> 21 ggcctccgct ctcgcttcgc ct 22 <210> 22 <211> 19 <212> DNA <213> Homo sapiens <400> 22 tcgtttctgc tcccggagc 19 <210> 23 <211> 19 <212> DNA <213> Homo sapiens <400> 23 tcgtttctgc gcccggagc 19 <210> 24 <211> 19 <212> DNA <213> Homo sapiens <400> 24 tcgtttctgc ccccggagc 19 <210> 25 <211> 19 <212> DNA <213> Homo sapiens <400> 25 tcgtttctgc acccggagc 19 <210> 26 <211> 20 <212> DNA <213> Homo sapiens <400> 26 cttgtctcgt agctgcagcc 20 <210> 27 <211> 15 <212> PNA <213> Homo sapiens <400> 27 gtttctgctc ccgga 15 <210> 28 <211> 15 <212> PNA <213> Homo sapiens <400> 28 gtttctgcgc ccgga 15 <210> 29 <211> 15 <212> PNA <213> Homo sapiens <400> 29 gtttctgccc ccgga 15 <210> 30 <211> 15 <212> PNA <213> Homo sapiens <400> 30 gtttctgcac ccgga 15 <210> 31 <211> 15 <212> PNA <213> Homo sapiens <400> 31 tgctgtcgat cgcac 15 <210> 32 <211> 15 <212> PNA <213> Hepatitis C Virus <400> 32 cttggggata ggctg 15 <210> 33 <211> 15 <212> PNA <213> Hepatitis C Virus <400> 33 ccatcctgcc caccc 15 <210> 34 <211> 15 <212> PNA <213> Hepatitis C Virus <400> 34 ccatgaaggg cgaga 15 <210> 35 <211> 15 <212> PNA <213> Hepatitis C Virus <400> 35 ctcgtttccg tacag 15 <210> 36 <211> 15 <212> PNA <213> Hepatitis C Virus <400> 36 gagcccagga ccggt 15 <210> 37 <211> 581 <212> DNA <213> Homo sapiens <400> 37 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgt 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcncccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgc gac gggcg gacgggcg gacgggcg ccagcgcg cc ggggcg cc <210> 38 <211> 581 <212> DNA <213> Homo sapiens <400> 38 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgg 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcncccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgc gac gggcg gacgggcg gacgggcg gacgggcg ccgcgc cc <210> 39 <211> 581 <212> DNA <213> Homo sapiens <400> 39 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctga 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcncccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcggggtg aaagaggcgg gacgggcg gacgggcg gacgggcg gacgggccag cc <210> 40 <211> 581 <212> DNA <213> Homo sapiens <400> 40 atgagccaga ctccagggag gcctagaagt gggcaagggg aaacgggaaa ggaggaagat 60 ggtatgggtg tgcctggtta ggggtgggag tgctggacgg agttcgggac aagaggggct 120 ctgcagccat tggcacacaa tgcctgggag tccctgctgg tgctgggatc atcccagtga 180 gccctgggag ggaactgaag acccccaatt accaatgcat ctgttttcaa aaccgacggg 240 gggaaggaca tgcctaggtt caaggatacg tgcaggcttg gatgactccg ggccattagg 300 gagcctccgg agcaccttga tcctcagacg ggcctgatga aacgagcatc tgattcagca 360 ggcctgggtt cgggcccgag aacctgcgtc tcccgcgagt tcccgcgagg caagtgctgc 420 aggtgcgggg ccaggagcta ggtttcgttt ctgcncccgg agccgccctc agcacagggt 480 ctgtgagttt catttcttcg ccggcgcggg gcggggctgg gcgcgggggag aaagaggcgg aaagaggcgc gacagggcg gagagggcg gg aagaggccag 540g <210> 41 <211> 1802 <212> DNA <213> Homo sapiens <400> 41 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacggaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 42 <211> 1802 <212> DNA <213> Homo sapiens <400> 42 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccaccgaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 43 <211> 1802 <212> DNA <213> Homo sapiens <400> 43 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacagaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 44 <211> 1802 <212> DNA <213> Homo sapiens <400> 44 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccactgaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 45 <211> 1802 <212> DNA <213> Homo sapiens <400> 45 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatggcacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 46 <211> 1802 <212> DNA <213> Homo sapiens <400> 46 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgacacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 47 <211> 1802 <212> DNA <213> Homo sapiens <400> 47 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgccacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 48 <211> 1802 <212> DNA <213> Homo sapiens <400> 48 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgtcacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 49 <211> 1802 <212> DNA <213> Homo sapiens <400> 49 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggcgt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 50 <211> 1802 <212> DNA <213> Homo sapiens <400> 50 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggtgt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 51 <211> 1802 <212> DNA <213> Homo sapiens <400> 51 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggagt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 52 <211> 1802 <212> DNA <213> Homo sapiens <400> 52 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggggt gatgncacca aggnagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 53 <211> 1802 <212> DNA <213> Homo sapiens <400> 53 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca agggagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 54 <211> 1802 <212> DNA <213> Homo sapiens <400> 54 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggaagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 55 <211> 1802 <212> DNA <213> Homo sapiens <400> 55 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggtagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 56 <211> 1802 <212> DNA <213> Homo sapiens <400> 56 gaattcctgc cagaaagtag agaggtattt agcactctgc cagggccaac gtagtaagaa 60 atttccagag aaaatgctta cccaggcaag cctgtntaaa acaccaaggg gaagcaaact 120 ccagttaatt ctgggctggg ttggtgacta aggttgaggt tgatctgagg ttgagacctt 180 cctctttgga tcaccagctt tcagctcagg gcctgccaat gagtaaatga tagttaacag 240 gtcctggagg ggaatcagct gcccagatac aaagatggga ttcaggtggc agatggaccc 300 gaagaggaca tggagagaaa gaggaagctc ctacagacac ctgggtttcc actcattctc 360 attccctaag ctaacaggca taagccagct ggcaatgcac ggtcccattt gttctcactg 420 ccacngaaag catgtttata gtcttccagc agcaacgcca ggtgtctagg cacagatgaa 480 cccctcctta ggatccccac tgctcatcat agtgcctacc tttgttaaag tactagtcac 540 gcagtgtcac aaggaatgtt tacttttcca aatccccagc tagaggccag ggatgggtca 600 tctatttcta tatagcctgc acccagattg taggacagag ggcatgctng gtaaatatgt 660 gttcattaac tgagattaac cttccctgag ttttctcaca ccaaggtgag gaccatgtcc 720 ctgtttccat cactccctct ccttctcctg agtatggtgg cagcgtctta ctcagaaact 780 gtgacctgtg aggatgccca aaagacctgc cctgcagtga ttgcctgtag ctctccaggc 840 atcaacggct t cccaggcaa agatgggngt gatgncacca aggcagaaaa gggggaacca 900 ggtacgtgtt gggctgttct gtctctgcaa ttctttacct tccagaggaa actgcctggg 960 gatatgagga gactgatgtc ctatttgagt atatttttct caactatact gtaactcaaa 1020 acagagattc agctcgaatt ccacacagca gtttgtgact aatagttgtc ttgccagccc 1080 aggaaagtgg cccacaggtc aggccatccc gtgggacaca ggatgaattt ttcttctctg 1140 ggtcattgtc atgtcagacc cctattcact tcagtaggga tggcaccagg ttcaagaggc 1200 caaagaagag atggagtcag caaacaaaca taggttttac tgggggaatc tgtttacagg 1260 gagatccagc agcagtgggc tggacaggag aacaacaact actggtaaaa acaaatgcag 1320 ttaattttca ctttgcaccc tccctgcagc aacctccacg tggcaacttt atttcttaag 1380 ttattgctct caggtgcaca ccatacagtt attgagagca gtgctcagaa aggtcagtcc 1440 tgggtcaagg tctcccttct cctgagaagg gattgggcat caaactcttg aagagagaga 1500 gcaagaacat agatattaag tcacatttcc tttgtcttcc aacaggccaa gggctcagag 1560 gcttacaggg cccccctgga aagttggggc ctccaggaaa tccagggcct tctgggtcac 1620 caggaccaaa gggccaaaaa ggagaccctg gaaaaagtcc gggtaaggac cccagcaagg 1680 tctgagctga cttcaccca g ggttctgaga ccttgagtat ctggtaagag gtgccccttc 1740 tcctgttcct tcaaaggaag atacccaaat ttgctttctg acccagtgcc ctcagccctc 1800 tc 1802 <210> 57 <211> 18 <212> DNA <213> Homo sapiens <400> 57 ngtgatggca ccaaggna 18 <210> 58 <211> 18 <212> DNA <213> Homo sapiens <400> 58 ngtgatgaca ccaaggna 18 <210> 59 <211> 18 <212> DNA <213> Homo sapiens <400> 59 ngtgatgtca ccaaggna 18 <210> 60 <211> 18 <212> DNA <213> Homo sapiens <400> 60 ngtgatgcca ccaaggna 18 <210> 61 <211> 18 <212> DNA <213> Homo sapiens <400> 61 ggtgatgnca ccaaggna 18 <210> 62 <211> 18 <212> DNA <213> Homo sapiens <400> 62 agtgatgnca ccaaggna 18 <210> 63 <211> 18 <212> DNA <213> Homo sapiens <400> 63 tgtgatgnca ccaaggna 18 <210> 64 <211> 18 <212> DNA <213> Homo sapiens <400> 64 cgtgatgnca ccaaggna 18 <210> 65 <211> 18 <212> DNA <213> Homo sapiens <400> 65 ngtgatgnca ccaaggga 18 <210> 66 <211> 18 <212> DNA <213> Homo sapiens <400> 66 ngtgatgnca ccaaggaa 18 <210> 67 <211> 18 <212> DNA <213> Homo sapiens <400> 67 ngtgatgnca ccaaggta 18 <210> 68 <211> 18 <212> DNA <213> Homo sapiens <400> 68 ngtgatgnca ccaaggca 18 <210> 69 <211> 30 <212> DNA <213> Homo sapiens <400> 69 acacacccgt ttccaccctg gagaggccag 30 <210> 70 <211> 30 <212> DNA <213> Homo sapiens <400> 70 tgcgcagtgc tggagtgcgg cctccgctct 30 <210> 71 <211> 19 <212> DNA <213> Homo sapiens <400> 71 cctgtgagga actactgtc 19 <210> 72 <211> 20 <212> DNA <213> Homo sapiens <400> 72 ggtgcacggt ctacgagacc 20

[Brief description of the drawings]

FIG. 1 is a schematic diagram of a DNA chip according to the present embodiment.

FIG. 2 is a schematic diagram of a PNA chip according to the embodiment.

FIG. 3 is a flowchart illustrating a method according to the embodiment.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... DNA probe 2 ... DNA probe 3 ... DNA probe 4 ... DNA probe 5 ... DNA probe 6 ... Substrate 7 ... PNA probe 8 ... PNA probe 9 ...・ PNA probe 10 ・ ・ ・ PNA probe 11 ・ ・ ・ PNA probe 12 ・ ・ ・ Base 13 ・ ・ ・ Gold electrode 14 ・ ・ ・ Connection

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 33/566 G01N 33/576 Z 33/569 C12R 1:93 33/576 C12N 15/00 ZNAA // ( C12Q 1 / 68F C12R 1:93) (72) Inventor Shunji Midai 1-1-1, Shibaura, Minato-ku, Tokyo Inside Toshiba Corporation Head Office (72) Inventor Hirohiko Ota 1-1-1, Shibaura, Minato-ku, Tokyo No. 1 F-term in Toshiba head office (reference) 4B024 AA01 AA11 CA02 CA09 CA11 HA14 4B029 AA07 BB13 BB20 CC08 CC10 CC13 FA01 4B063 QA01 QA12 QA18 QA19 QQ10 QQ42 QR32 QR55 QR83 QR84 QS34 QS36 QX02 QX04

Claims (33)

[Claims] 1. A method for obtaining first information about nucleic acids of an individual exposed to a specific disease and second information about nucleic acids from pathogenic microorganisms present in said individual, wherein said pathogenic microorganisms are A method relating to a specific disease, comprising: (a) reacting an extract of nucleic acid from the individual with a probe-immobilized substrate comprising a first probe and a second probe; Wherein the first probe detects the presence of a specific nucleic acid sequence of the pathogenic microorganism, the pathogenic microorganism is associated with the specific disease, and the second probe detects the presence of a specific nucleic acid of the individual. And (b) obtaining the first information by detecting the presence of a nucleic acid bound to the first probe as a result of the reaction of (a); Nucleus bound to Obtaining second information by detecting the presence of. 2. The nucleic acid of the individual is associated with responsiveness to the treatment of the disease, and the presence of both the nucleic acid from the pathogenic microorganism and the nucleic acid of the individual correlates with the responsiveness to the treatment of the disease. The method of claim 1, wherein 3. The method of claim 1, further comprising, prior to the reaction of (a), amplifying the extract of a nucleic acid from the individual, and subjecting the extract to obtaining an amplified nucleic acid. 4. The method of claim 1, wherein said individual is a human. 5. The method of claim 1, wherein said extract of nucleic acid is prepared from whole blood. 6. The method of claim 5, wherein said nucleic acid from said individual is a human genomic nucleic acid and said nucleic acid from said pathogenic microorganism is a genomic nucleic acid. 7. The nucleic acid from the individual is a human genomic nucleic acid, and the nucleic acid from the pathogenic microorganism is RNA;
2. The method according to claim 1, wherein a reverse transcription reaction is performed prior to the reaction (a). 8. The method according to claim 3, wherein the nucleic acid from the individual is a human genomic nucleic acid, the nucleic acid from the pathogenic microorganism is RNA, and a reverse transcription reaction is performed prior to the amplification. 9. The method according to claim 6, wherein the nucleic acid extract is obtained using a kit for extracting human genome. 10. The kit for extracting human genome according to claim 1, wherein the kit for extracting human genome is QIA.
The method according to claim 9, which is an amp DNA Blood Midi Kid. 11. The method according to claim 1, wherein the specific disease is hepatitis, and the pathogenic microorganism is hepatitis virus. 12. The method according to claim 3, wherein the disease is hepatitis and the pathogenic microorganism is hepatitis virus. 13. The first probe detects hepatitis C virus genome and the second probe detects MxA
The method according to claim 11, wherein the SNP in the promoter region is detected. 14. The method according to claim 11, wherein the first probe detects a hepatitis C virus genome and the second probe detects a polymorphism in the MBL gene. 15. The first probe is provided with at least one kind, and at least one kind includes a sequence selected from the following: (a) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5 A base sequence represented by a sequence selected from the group consisting of: (b) a base sequence represented by a sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7,
(C) a complementary sequence of a base sequence selected from (a) and (b); and the second probe is provided with at least one kind, and at least one kind includes a sequence selected from the following; The base sequence shown in SEQ ID NO: 16 in the following sequence listing, (bt455) several bases except for the base at position 455 in the base sequence shown in (at455) are deleted, substituted, or one or more bases. An added modified nucleotide sequence, (ct455) a nucleotide sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 16, (dt455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 16, (et455) (At455) a complementary sequence of a base sequence selected from (dt455), (ag455) the base sequence of SEQ ID NO: 17 in the following sequence listing, (bg455) position 455 in the base sequence of (ag455) A modified base sequence in which several bases except for the base are deleted, substituted, or one or more bases are added, (cg455) (Dg455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 17, (eg455) selected from the (ag455) to (dg455) (Aa455) a base sequence represented by SEQ ID NO: 18 in the following sequence listing; (ba455) several bases excluding the base at position 455 in the base sequence represented by (aa455) are deleted. (Ca455) a nucleotide sequence comprising the sequence shown at positions 441 to 455 of SEQ ID NO: 18, (da455) at positions 449 to 459 of SEQ ID NO: 18 (Ea455) a complementary sequence of a base sequence selected from (aa455) to (da455), (ac455) a base sequence represented by SEQ ID NO: 19 in the following sequence listing, (bc455) (ac455) a modified base sequence in which several bases except for the base at position 455 in the base sequence shown in SEQ ID NO: 455 have been deleted, substituted, or added with one or more bases, (cc (455) a nucleotide sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 19, (dc455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 19, (ec455) the (ac455) to (dc455) (Aa420) a base sequence represented by SEQ ID NO: 37 in the following sequence list, (ba420) several bases excluding the base at position 420 in the base sequence represented by (aa420) Has been deleted, substituted, or modified base sequence to which one or more bases have been added, (ca420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 37, (da420) the (aa420) to (ca420) (Ac420) a base sequence represented by SEQ ID NO: 38 in the following sequence listing, (bc420) several nucleotides excluding the base at position 420 in the base sequence represented by (ac420). Bases are deleted, substituted, or modified base sequence to which one or more bases are added, (cc420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 38, (dc420) a complementary sequence of a base sequence selected from the (ac420) to (cc420), (at420) a base sequence represented by SEQ ID NO: 39 in the following sequence listing, (bt420) a base sequence represented by the (at420) A modified base sequence in which several bases except for the base at position 420 are deleted, substituted, or added with one or more bases, (ct420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 39 (Dt420) a complementary sequence of a base sequence selected from (at420) to (ct420) (ag420) a base sequence represented by SEQ ID NO: 40 in the following sequence listing, (bg420) a base sequence represented by (at420) A modified base sequence in which several bases except for the base at position 420 are deleted, substituted, or added with one or more bases, (cg420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 40 (Dg420) a complementary sequence of a base sequence selected from the (ag420) to (cg420), (ag221) a base sequence represented by SEQ ID NO: 41 in the following sequence list, (bg221) (Ag221) a modified base sequence in which several bases except the base at position 425 in the base sequence shown in (ag221) are deleted, substituted, or added with one or more bases, (cg221) 418 to 418 of SEQ ID NO: 41 A nucleotide sequence comprising the sequence shown at position 432, (dg221) a nucleotide sequence comprising the sequence shown at positions 421 to 430 of SEQ ID NO: 41 (eg221) a complementary sequence of a nucleotide sequence selected from the (ag221) to (dg221) (Ac221) a base sequence represented by SEQ ID NO: 42 in the following sequence listing, (bc221) several bases excluding the base at position 425 in the base sequence represented by (ac221) are deleted, substituted, or 1 A modified base sequence to which the above bases are added, (cc221) a base sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 42, (dc221) a base sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 42 (ec221) a complementary sequence of the base sequence selected from (ac221) to (dc221), (aa221) a base sequence represented by SEQ ID NO: 43 in the following sequence listing, (ba221) (aa221) a modified base sequence in which several bases except for the base at position 425 in the base sequence shown in (aa221) are deleted, substituted, or added with one or more bases, (ca221) 418 to 432 of SEQ ID NO: 43 A nucleotide sequence containing the sequence shown at position (da221) a nucleotide sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 43 (ea221) a complementary sequence of a nucleotide sequence selected from the (aa221) to (da221), (at221) the base sequence of SEQ ID NO: 44 in the following sequence listing, (bt221) several bases except for the base at position 425 in the base sequence shown in (at221) are deleted, substituted, or one or more (Ct221) a nucleotide sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 44, (dt221) a nucleotide sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 44 ( et221) a complementary sequence of a base sequence selected from the (at221) to (dt221), (ag54) a base sequence represented by SEQ ID NO: 45 in the following sequence listing, (bg54) the (a g54) a modified base sequence in which several bases except for the base at position 875 in the base sequence shown in SEQ ID NO: 45 are deleted, substituted, or added with one or more bases, (cg54) SEQ ID NO: 45-874 to 876 (Dg54) a nucleotide sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 45 (eg54) a complementary sequence of a nucleotide sequence selected from the (ag54) to (dg54), aa54) the base sequence of SEQ ID NO: 46 in the following sequence listing, (ba54) several bases except for the base at position 875 in the base sequence shown in (aa54) are deleted, substituted, or one or more A modified base sequence to which bases are added, (ca54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 46, (da54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 46 (ea54 ) The complementary sequence of the base sequence selected from the (aa54) to (da54), (ac54) the base sequence of SEQ ID NO: 47 in the following sequence listing, (bc54) the base sequence of (ac54) A modified base sequence in which several bases except the base at position 875 in the base sequence have been deleted, substituted, or added with one or more bases, (cc54) a sequence shown at positions 874 to 876 of SEQ ID NO: 47. (Dc54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 47 (ec54) a complementary sequence of a base sequence selected from the above (ac54) to (dc54), (at54) The base sequence shown in SEQ ID NO: 47, (bt54) A few bases except the base at position 875 in the base sequence shown in (at54) were deleted, substituted, or one or more bases were added. Modified nucleotide sequence, (ct54) a nucleotide sequence comprising the sequence shown at positions 874 to 876 of SEQ ID NO: 48, (dt54) a nucleotide sequence comprising the sequence shown at positions 869 to 880 of SEQ ID NO: 48 (et54) ~ (Dt54), a complementary sequence of a base sequence selected from (ag52-57) a base sequence represented by SEQ ID NO: 45 in the following sequence listing, (bg52-57) a base represented by (ag52-57) A modified base sequence in which several bases except for the base at position 875 in the column are deleted, substituted, or added with one or more bases, (cg52-57) a sequence represented by positions 868 to 885 of SEQ ID NO: 45 (Dg52-57) a complementary sequence of the base sequence selected from (ag52-57) to (cg52-57), (aa52-57) a base sequence represented by SEQ ID NO: 46 in the following sequence list, (ba52-57) a modified base sequence in which several bases except the base at position 875 in the base sequence shown in (aa52-57) are deleted, substituted, or added with one or more bases, (ca52 -57) a base sequence comprising the sequence shown at positions 886 to 885 of SEQ ID NO: 46, (da52-57) a complementary sequence of the base sequence selected from (aa52-57) to (ca52-57), (ac52- 57) the base sequence of SEQ ID NO: 47 in the following sequence listing, (bc52-57) several bases except the base at position 875 in the base sequence shown in (ac52-57) are deleted and substituted, Or a modified base sequence to which one or more bases have been added (Cc52-57) a base sequence comprising the sequence shown at positions 886 to 885 of SEQ ID NO: 47, (dc52-57) a complementary sequence of the base sequence selected from (ac52-57) to (cc52-57), (at52-57) a base sequence represented by SEQ ID NO: 47 in the following sequence listing, (bt52-57) several bases except for the base at position 875 in the base sequence represented by (at52-57) are deleted, Substituted or modified base sequence to which one or more bases have been added, (ct52-57) a base sequence containing the sequence shown at positions 886 to 885 of SEQ ID NO: 48, (dt52-57) the (at52-57) to The method according to claim 11, wherein a sequence complementary to a base sequence selected from (ct52-57); 16. The first probe comprises at least one kind, and at least one kind contains a sequence selected from the following: (a) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5 A base sequence represented by a sequence selected from the group consisting of: (b) a base sequence represented by a sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7,
(C) a complementary strand of a base sequence selected from (a) and (b); and the second probe is provided with one or more kinds,
(At455) a base sequence represented by SEQ ID NO: 16 in the following sequence listing; (bt455) a base at position 455 in the base sequence represented by (at455); (Ct455) a base sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 16, a modified base sequence in which a few bases have been deleted, substituted, or added with one or more bases, (dt455) SEQ ID NO: 16, a base sequence comprising the sequence shown at positions 449 to 459, (et455) a complementary sequence of the base sequence selected from (at455) to (dt455), (ag455) a base shown in SEQ ID NO: 17 in the following sequence list. (Bg455) a modified base sequence in which several bases except for the base at position 455 in the base sequence shown in (ag455) are deleted, substituted, or added with one or more bases, (cg455) sequence A base sequence comprising the sequence shown at positions 441-455 of SEQ ID NO: 17, (dg455) a base sequence comprising the sequence shown at positions 449-459 of SEQ ID NO: 17 (Eg455) a complementary sequence of a base sequence selected from the (ag455) to (dg455), (aa455) a base sequence represented by SEQ ID NO: 18 in the following sequence listing, (ba455) a base sequence represented by the (aa455) A modified base sequence in which several bases except for the base at position 455 are deleted, substituted, or added with one or more bases, (ca455) a base including the sequence shown at positions 441 to 455 of SEQ ID NO: 18. Sequence, (da455) a base sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 18, (ea455) a complementary sequence of a base sequence selected from the (aa455) to (da455), (ac455) A base sequence represented by SEQ ID NO: 19, (bc455) a modification in which several bases except for the base at position 455 in the base sequence represented by (ac455) are deleted, substituted, or added with one or more bases; Base sequence, (cc455) a base sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 19, (dc455) a base containing the sequence shown at positions 449 to 459 of SEQ ID NO: 19 Sequence, (ec455) a complementary sequence of a base sequence selected from (ac455) to (dc455), (aa420) a base sequence represented by SEQ ID NO: 37 in the following sequence listing, (ba420) a base represented by (aa420) A modified base sequence in which several bases except for the base at position 420 in the sequence are deleted, substituted, or added with one or more bases, (ca420) including the sequence shown at positions 415 to 425 of SEQ ID NO: 37 Base sequence, (da420) a complementary sequence of the base sequence selected from (aa420) to (ca420), (ac420) a base sequence represented by SEQ ID NO: 38 in the following sequence listing, (bc420) represented by (ac420) A modified base sequence in which several bases excluding the base at position 420 in the base sequence have been deleted, substituted, or added with one or more bases, (cc420) a sequence shown at positions 415 to 425 of SEQ ID NO: 38. (Dc420) a complementary sequence of the base sequence selected from (ac420) to (cc420), (at420) a base sequence represented by SEQ ID NO: 39 in the following sequence listing. (bt420) a modified base sequence in which several bases except the base at position 420 in the base sequence shown in (at420) are deleted, substituted, or added with one or more bases, (ct420) SEQ ID NO: 39 (Dt420) a complementary sequence of the base sequence selected from (at420) to (ct420), (ag420) a base sequence represented by SEQ ID NO: 40 in the following sequence table. (Bg420) a modified base sequence in which several bases except the base at position 420 in the base sequence shown in (at420) are deleted, substituted, or added with one or more bases, (cg420) SEQ ID NO: (Dg420) a complementary sequence of a base sequence selected from (ag420) to (cg420), (ag221) a base sequence represented by SEQ ID NO: 41 in the following sequence list, Sequence, (bg221) several bases except for the base at position 425 in the base sequence shown in (ag221) were deleted, substituted, or one or more bases were added Modified nucleotide sequence, (cg221) a nucleotide sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 41, (dg221) a nucleotide sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 41 (eg221) the (ag221) ~ (Dg221) a complementary sequence of a base sequence selected from: (ac221) the base sequence shown in SEQ ID NO: 42 of the following sequence listing, (bc221) excluding the base at position 425 in the base sequence shown in (ac221) A modified base sequence in which several bases have been deleted, substituted or added with one or more bases, (cc221) a base sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 42, (dc221) SEQ ID NO: 42 A base sequence comprising the sequence shown at positions 421 to 430 of (ec221) a complementary sequence of a base sequence selected from the (ac221) to (dc221), (aa221) a base sequence represented by SEQ ID NO: 43 in the following sequence table, (ba221) Several nucleotides except for the nucleotide at position 425 in the nucleotide sequence shown in (aa221) were deleted, substituted, or one or more nucleotides were added. A decorative base sequence, (ca221) a base sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 43, (da221) a base sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 43 (ea221) the (aa221) ~ (Da221) the complementary sequence of the base sequence selected from (at221) the base sequence shown in SEQ ID NO: 44 of the following sequence listing, (bt221) excluding the base at position 425 in the base sequence shown in the above (at221) A modified base sequence in which several bases are deleted, substituted, or added with one or more bases, (ct221) a base sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 44, (dt221) SEQ ID NO: 44 A nucleotide sequence comprising the sequence shown at positions 421 to 430 of (et221) a complementary sequence of a nucleotide sequence selected from the (at221) to (dt221), (ag54) a nucleotide sequence represented by SEQ ID NO: 45 in the following sequence table, (bg54) a modification in which several bases except the base at position 875 in the base sequence shown in (ag54) are deleted, substituted, or one or more bases are added (Cg54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 45, (dg54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 45 (eg54) the above (ag54) to (dg54) a complementary sequence of a base sequence selected from (aa54) a base sequence represented by SEQ ID NO: 46 in the following sequence listing, and (ba54) a number excluding the base at position 875 in the base sequence represented by (aa54). (Ca54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 46, (da54) a modified base sequence in which one or more bases have been deleted, substituted, or added with one or more bases, Nucleotide sequence containing the sequence shown at positions 869 to 880 (ea54) The complementary sequence of the nucleotide sequence selected from (aa54) to (da54), (ac54) the nucleotide sequence represented by SEQ ID NO: 47 in the following sequence listing, (bc54) a modified base sequence in which several bases except the base at position 875 in the base sequence shown in (ac54) have been deleted, substituted, or added with one or more bases, (cc 54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 47, (dc54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 47 (ec54) from the (ac54) to (dc54) The complementary sequence of the selected base sequence, (at54) the base sequence shown in SEQ ID NO: 47 of the following sequence listing, (bt54) several bases excluding the base at position 875 in the base sequence shown in (at54), A modified base sequence to which deletion, substitution, or addition of one or more bases, (ct54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 48, (dt54) positions 869 to 880 of SEQ ID NO: 48 (Et54) a complementary sequence of a base sequence selected from (at54) to (dt54), (ag52-57) a base sequence represented by SEQ ID NO: 45 in the following sequence listing, (bg52- 57) a modified base sequence in which several bases except the base at position 875 in the base sequence shown in (ag52-57) are deleted, substituted, or one or more bases are added, (cg52-57) A nucleotide sequence comprising the sequence shown at positions 868 to 885 of SEQ ID NO: 45, (dg52-57) a complementary sequence of the nucleotide sequence selected from (ag52-57) to (cg52-57), (aa52-57) Nucleotide sequence shown in SEQ ID NO: 46 of the sequence listing, (ba52-57) Several nucleotides excluding the base at position 875 in the nucleotide sequence shown in (aa52-57) are deleted, substituted, or one or more. (Ca52-57) a base sequence containing the sequence shown at positions 886 to 885 of SEQ ID NO: 46, (da52-57) the (aa52-57) to (ca52-57) The complementary sequence of the selected base sequence, (ac52-57) the base sequence shown in SEQ ID NO: 47 of the following sequence listing, (bc52-57) the base at position 875 in the base sequence shown in (ac52-57). A modified base sequence in which several bases except for deletion, substitution, or addition of one or more bases, (cc52-57) a base sequence including the sequence shown at positions 886 to 885 of SEQ ID NO: 47, (dc52- 57) A salt selected from (ac52-57) to (cc52-57) The complementary sequence of the base sequence, (at52-57) the base sequence shown in SEQ ID NO: 47 of the following sequence listing, (bt52-57) several bases excluding the base at position 875 in the base sequence shown in (at52-57) Base sequence is deleted, substituted, or a modified base sequence to which one or more bases are added, (ct52-57) a base sequence containing the sequence shown at positions 886 to 885 of SEQ ID NO: 48, (dt52-57) 13. The method according to claim 12, wherein a complementary sequence of a base sequence selected from (at52-57) to (ct52-57). 17. The method according to claim 17, wherein the amplification comprises a sense strand represented by at least one base sequence selected from the group consisting of SEQ ID NO: 8 and SEQ ID NO: 9, The method according to claim 12, wherein the method is performed by PCR using an antisense strand represented by a base sequence selected at least one from the group of SEQ ID NO: 14. 18. A method for detecting the presence of a substrate and a particular nucleic acid sequence of a pathogenic microorganism,
A first probe immobilized on the substrate, wherein the microorganism is associated with a specific disease and is different from the first probe immobilized on the substrate for detecting the presence of a specific nucleic acid in an individual A second probe having a base sequence, wherein the nucleic acid of the individual is
A probe-immobilized substrate, which is related to responsiveness to treatment of the disease. 19. The method according to claim 18, wherein the pathogenic microorganism is hepatitis C virus, and the drug used in the treatment is IFN.
4. The probe-immobilized substrate according to 1. 20. The probe-immobilized substrate according to claim 18, wherein the nucleic acid of the individual is an MxA promoter region. 21. The probe-immobilized substrate according to claim 18, wherein the nucleic acid of the individual is a gene encoding MBL. 22. The probe-immobilized substrate according to claim 18, wherein the first probe detects a hepatitis C virus genotype. 23. A base, which is immobilized on the base and has the following sequence: (a) a base represented by a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5 Sequence, (b) a base sequence represented by a sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7,
(C) a complementary sequence of a base sequence selected from (a) and (b); a first probe including a sequence selected from at least one of the following: immobilized on the substrate and having the following sequence; The nucleotide sequence represented by SEQ ID NO: 16 in the sequence listing, (bt455) Several nucleotides except for the nucleotide at position 455 in the nucleotide sequence represented by (at455) are deleted or substituted, or one or more nucleotides are added. Modified nucleotide sequence, (ct455) a nucleotide sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 16, (dt455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 16, (et455) (at455) to a complementary sequence of a base sequence selected from (dt455), (ag455) a base sequence represented by SEQ ID NO: 17 in the following sequence listing, (bg455) position 455 in the base sequence represented by (ag455). A modified base sequence in which several bases excluding bases are deleted, substituted, or one or more bases are added, (cg455) SEQ ID NO: 17 (Dg455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 17, (eg455) a nucleotide sequence selected from the (ag455) to (dg455) Complementary sequence of the sequence (aa455) The base sequence shown in SEQ ID NO: 18 in the following sequence listing, (ba455) Several bases except for the base at position 455 in the base sequence shown in (aa455) are deleted or substituted. Or a modified base sequence to which one or more bases have been added, (ca455) a base sequence containing the sequence shown at positions 441 to 455 of SEQ ID NO: 18, (da455) a sequence shown at positions 449 to 459 of SEQ ID NO: 18. (Ea455) a complementary sequence of a base sequence selected from the (aa455) to (da455) (ac455) a base sequence represented by SEQ ID NO: 19 in the following sequence listing, (bc455) represented by (ac455) A modified base sequence in which several bases except for the base at position 455 in the base sequence have been deleted or substituted, or one or more bases have been added, (cc455) SEQ ID NO: 19 (Dc455) a nucleotide sequence containing the sequence shown at positions 449 to 459 of SEQ ID NO: 19, (ec455) a nucleotide sequence selected from the (ac455) to (dc455) (Aa420) a base sequence represented by SEQ ID NO: 37 in the following sequence listing, (ba420) several bases except for the base at position 420 in the base sequence represented by (aa420) are deleted or substituted. Or a modified base sequence to which one or more bases are added, (ca420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 37, (da420) a base selected from the (aa420) to (ca420) Complementary sequence of the sequence (ac420) The base sequence shown in SEQ ID NO: 38 of the following sequence listing, (bc420) Several bases except for the base at position 420 in the base sequence shown in (ac420) are deleted and substituted. Or a modified base sequence to which one or more bases have been added, (cc420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 38, (dc420) the (ac420) The complementary sequence of the base sequence selected from (cc420) (at420) the base sequence shown in SEQ ID NO: 39 in the following sequence listing, (bt420) several nucleotides excluding the base at position 420 in the base sequence shown in (at420) Base sequence is deleted, substituted, or modified base sequence to which one or more bases have been added, (ct420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 39, (dt420) the (at420) to The complementary sequence of the base sequence selected from (ct420) (ag420) the base sequence shown in SEQ ID NO: 40 of the following sequence listing, (bg420) several nucleotides excluding the base at position 420 in the base sequence shown in (at420) (Cg420) a base sequence containing the sequence shown at positions 415 to 425 of SEQ ID NO: 40, (dg420) the (ag420) to (cg420) a complementary sequence of a base sequence selected from: (ag221) a base sequence represented by SEQ ID NO: 41 in the following sequence list, (bg221) represented by (ag221) A modified base sequence in which several bases except for the base at position 425 in the base sequence have been deleted, substituted, or added with one or more bases, (cg221) a sequence shown at positions 418 to 432 of SEQ ID NO: 41. (Dg221) a base sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 41 (eg221) a complementary sequence of a base sequence selected from the (ag221) to (dg221), (ac221) The base sequence shown in SEQ ID NO: 42, (bc221) Several bases except the base at position 425 in the base sequence shown in (ac221) were deleted, substituted, or one or more bases were added. Modified nucleotide sequence, (cc221) a nucleotide sequence containing the sequence shown at positions 418 to 432 of SEQ ID NO: 42, (dc221) a nucleotide sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 42 (ec221) the (ac221) ~ (Dc221) a complementary sequence of a base sequence selected from: (aa221) a base sequence represented by SEQ ID NO: 43 in the following sequence listing, (ba221) a salt represented by (aa221) A modified base sequence in which several bases except the base at position 425 in the base sequence have been deleted, substituted, or added with one or more bases, (ca221) a sequence shown at positions 418 to 432 of SEQ ID NO: 43. (Da221) a base sequence containing the sequence shown at positions 421 to 430 of SEQ ID NO: 43 (ea221) a complementary sequence of a base sequence selected from the (aa221) to (da221), (at221) The nucleotide sequence shown in SEQ ID NO: 44, (bt221) Several nucleotides except for the nucleotide at position 425 in the nucleotide sequence shown in (at221) were deleted, substituted, or one or more nucleotides were added. A modified nucleotide sequence, (ct221) a nucleotide sequence comprising the sequence shown at positions 418 to 432 of SEQ ID NO: 44, (dt221) a nucleotide sequence comprising the sequence shown at positions 421 to 430 of SEQ ID NO: 44 (et221) ~ (Dt221) the complementary sequence of the base sequence selected from (ag54) the base sequence shown in SEQ ID NO: 45 of the following sequence listing, (bg54) the base shown in the above (ag54) A modified base sequence in which several bases except for the base at position 875 in the column are deleted, substituted, or added with one or more bases, (cg54) including the sequence shown at positions 874 to 876 of SEQ ID NO: 45 Nucleotide sequence, (dg54) a nucleotide sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 45 (eg54) a complementary sequence of a nucleotide sequence selected from the (ag54) to (dg54), (aa54) The base sequence represented by SEQ ID NO: 46, (ba54) a modification in which several bases except the base at position 875 in the base sequence represented by (aa54) are deleted, substituted, or added with one or more bases (Ca54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 46, (da54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 46 (ea54) the (aa54) to (da54) a complementary sequence of a base sequence selected from: (ac54) a base sequence represented by SEQ ID NO: 47 in the following sequence listing, (bc54) position 875 in the base sequence represented by (ac54) A modified base sequence in which several bases excluding bases are deleted, substituted, or one or more bases are added, (cc54) a base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 47, (dc54) A base sequence comprising the sequence shown at positions 869 to 880 of SEQ ID NO: 47 (ec54) a complementary sequence of a base sequence selected from the above (ac54) to (dc54), (at54) shown in SEQ ID NO: 47 of the following sequence table Base sequence, (bt54) a modified base sequence in which several bases except the base at position 875 in the base sequence shown in (at54) are deleted, substituted, or added with one or more bases, (ct54) A base sequence containing the sequence shown at positions 874 to 876 of SEQ ID NO: 48; (dt54) a base sequence containing the sequence shown at positions 869 to 880 of SEQ ID NO: 48 (et54) selected from the above (at54) to (dt54) (Ag52-57) a base sequence represented by SEQ ID NO: 45 in the following sequence listing, (bg52-57) a base at position 875 in the base sequence represented by (ag52-57), (Cg52-57) a base sequence containing the sequence shown at positions 868 to 885 of SEQ ID NO: 45, wherein a modified base sequence in which several bases except for have been deleted, substituted, or added with one or more bases, (dg52- 57) a complementary sequence of a base sequence selected from the (ag52-57) to (cg52-57), (aa52-57) a base sequence represented by SEQ ID NO: 46 in the following sequence listing, (ba52-57) the (aa52-57) -57) a modified base sequence in which several bases except for the base at position 875 in the base sequence shown in (57) are deleted, substituted, or added with one or more bases, (ca52-57) 886 of SEQ ID NO: 46 (Da52-57) The complementary sequence of the base sequence selected from (aa52-57) to (ca52-57), (ac52-57) SEQ ID NO: Base sequence shown in 47, (bc52-57) Several bases except the base at position 875 in the base sequence shown in (ac52-57) are deleted, substituted, or one or more bases are added. Modified base sequence, (cc52-57) SEQ ID NO: 4 7, a base sequence containing the sequence shown at positions 886 to 885, (dc52-57) a complementary sequence of the base sequence selected from (ac52-57) to (cc52-57), (at52-57) The nucleotide sequence shown in SEQ ID NO: 47, (bt52-57) Several nucleotides except for the nucleotide at position 875 in the nucleotide sequence shown in (at52-57) are deleted, substituted, or one or more nucleotides. (Ct52-57) a base sequence containing the sequence shown at positions 886 to 885 of SEQ ID NO: 48, (dt52-57) selected from the (at52-57) to (ct52-57) A second probe comprising at least one sequence selected from the following: a complementary sequence of a base sequence; 24. The probe-immobilized substrate according to claim 18, wherein the detection of the presence of the specific nucleic acid is performed electrochemically. 25. The probe-immobilized substrate according to claim 23, wherein the presence of the specific nucleic acid is detected electrochemically. 26. A method for determining the responsiveness of an individual to treatment of a disease comprising: wherein the disease is associated with the presence of a pathogenic microorganism in the individual; (a) an extract of a nucleic acid from the individual A first nucleic acid probe that hybridizes to the nucleic acid of the pathogenic microorganism (where the pathogenic microorganism is associated with the disease) and a second nucleic acid probe that hybridizes to the target nucleic acid of the individual (wherein the individual And (b) the nucleic acid of the pathogenic microorganism that binds to the first nucleic acid probe, and the second nucleic acid probe. Detecting the target nucleic acid of the individual that binds to
The presence of both the nucleic acid of the pathogenic microorganism that binds to the nucleic acid probe and the target nucleic acid of the individual that binds to the second nucleic acid probe correlates with responsiveness to treatment of the disease). 27. The method according to claim 26, wherein the probe is immobilized on a substrate. 28. The method according to claim 2, further comprising amplifying the nucleic acid from the individual prior to the reaction of (a).
7. The method according to 6. 29. A method for determining the susceptibility of an individual to a disease comprising: (a) a first nucleic acid probe that hybridizes to an extract of a nucleic acid from said individual and a nucleic acid of a pathogenic microorganism associated with the disease. And reacting a second nucleic acid probe that hybridizes to a target nucleic acid of the individual associated with susceptibility to the disease; (b) the nucleic acid of the pathogenic microorganism that binds to the first nucleic acid probe; Detecting the nucleic acid of the individual that binds to the second nucleic acid probe (wherein the first
The presence of both the nucleic acid of the pathogenic microorganism that binds to the nucleic acid probe and the nucleic acid of the individual that binds to the second nucleic acid probe is associated with the susceptibility of the individual to the disease). 30. The method according to claim 29, wherein the probe is immobilized on a substrate. 31. The method according to claim 2, further comprising amplifying the nucleic acid from the individual prior to the reaction of (a).
9. The method according to 9. 32. A first probe for detecting the presence of a specific nucleic acid of a pathogenic microorganism associated with a disease, and a second probe for detecting the presence of a specific nucleic acid of an individual associated with responsiveness to treatment for the disease. A method for producing a probe-immobilized substrate, comprising: fixing the first probe and the second probe to the substrate. 33. The method according to claim 32, wherein the substrate is selected from the group consisting of a base plate, a porous body, a microtiter plate, beads, spherical substances, particulate substances, magnetic substances, and magnetic beads.