CN104846074A - Probe used for TORCH detection, gene chip and kit thereof - Google Patents
Probe used for TORCH detection, gene chip and kit thereof Download PDFInfo
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Abstract
The invention discloses a set of probes used for TORCH detection, and comprises a toxoplasma TOX probe which is shown in a SEQ ID NO. 1; a rubella virus RV probe which is shown in a SEQ ID NO.2; a cytomegalovirus CMV probe which is shown in a SEQ ID NO.3; a herpes simplex virus HSV I type probe which is shown in a SEQ ID NO.4; and a herpes simplex virus HSV II type probe which is shown in a SEQ ID NO.5. The invention also comprises a gene chip containing the probe and a kit thereof, visible light passing through a biochip can be amplified, signal resolution is high, and a signal can be shoot by a common digital camera or directly read by naked eyes. Compared with a traditional fluorescent label, the gene chip has good effect. Through specific primer and probe selection, by combining a gene chip technology, detection efficiency and accuracy degree are increased, and requirement of on-site detection for hospital and antenatal testing.
Description
Technical field
The present invention relates to biological technical field, be specifically related to one group of probe detected for TORCH, gene chip and test kit
Background technology
TORCH refers to cause congenital intrauterine infection and perinatal infection and causes the pathogenic agent of peri-natal infant deformity, it is the English name abbreviation of one group of pathogenic micro-organism, wherein T (Toxopasma) is toxoplasma gondii, R (Rubella.Virus) is rubella virus, C (Cytomegalo.Virus) is giant cells, and H (Herpes.Virus) is namely herpes simplex I/II type.
This group infected by microbes has common feature, and mother and baby can be caused to infect.Easily there is primary infection due to endocrine alteration and immunity degradation in pregnant woman, in pregnant woman's body of previous infection, potential virus is also easily activated and recurrent infection occurs.When viremia occurs pregnant woman, virus is propagated by placenta or birth canal and is infected fetus, causes premature labor, miscarriage, stillborn foetus or monster etc., and causes the infringement of the multiple system of newborn infant, multiple organ, cause the symptoms such as dysnoesia in various degree.Be in organogenetic period pregnancy three months embryos just especially, be now infected by the virus, division and the propagation of cell or T suppression cell can be destroyed.Organogenetic period, can disorganize and organ structure with postoperative infection virus, and can form persistent infection, continues toxin expelling, can cause corresponding pathology after birth.The infection influencer population quality of TORCH, has important relationship with prenatal and postnatal care, wherein:
Toxoplasma gondii (TOX):
The fetal anomaly that pregnant early stage arch insect infection causes mainly comprises: hydrocephalus, little deformity of brain, choroidoretinitis and brain calcium rubella virus (RV):
Change.Hematogenous infection can cause fetus multiple organ necrotic lesions, as hepatosplenomegaly, myocarditis and thrombocytopenia etc.
RV is mainly through respiratory infectious, fetus teratogenesis can be made after infection of pregnant women, virus forms congenital infection by placental infection fetus, be called congenital rubella syndrome (CRS), mainly congenital cataract, congenital heart disease and neural heariing loss, within 20 weeks, postoperative infection person is almost without impact.Rubella-infection occurs in the pregnancy period more early, and the teratogenesis of fetus is also more serious.
Cytomegalovirus (CMV):
Can cause intrauterine fetal growth retardation, little capitiform, encephalitis, retina arteries and veins film inflammation, jaundice, hepatosplenomegaly, hemolytic anemia etc. after infection, neonatal mortality is higher, and the cmv infection rate caused by perinatal period breast milk toxin expelling is 63%.
Hsv (HSVI, II type):
HSV hides usually at neuroganglion.During gestation, the physiological change of parent makes HSV activate, and pregnant early infection can destroy plumule face and cause miscarriage, though pregnant middle and advanced stage sends out monster less, fetus an d neonate can be caused to fall ill.
The congenital malformation of torch infection and fetus is clinical workers always and is engaged in the focus that prenatal and postnatal care related researcher pays close attention to, and is also the emphasis that people study to the accurate detection of pathogenic agent and correct diagnostic method.ELISA, molecular hybridization and PCR were once widely used in prenatal and postnatal care and reduced the natality of Disabled children in clinical detection, but these methods all exist respective shortcoming.
Round pcr has advantage that is highly sensitive and direct-detection pathogen gene, but needs multitube system corresponding for four kinds of pathogenic agent, there is operation inconvenience, to operational requirement high.ElISA, Gold standard are not all directly detect pathogenic agent, and there is certain window phase, may cause false negative.Adopt gene chip to detect, it is large that it detects flux, and on a chip, once property completes four kinds of pathogen detection, directly carries out for pathogen gene, and result is more accurate, has the advantages such as detection sensitivity is high, specificity good, and required sample size is few.
The detection of traditional biochip technology carries out detecting based on the fluorescence radiation of fluorescence labeling probe.Its signal is weak, has to pass through and excites, could be luminous, needs expensive Laser Scanning Equipment.In clinical detection application process, need investment to purchase Laser Scanning Equipment owing to detecting unit, on the one hand, significantly increase the cost of investment of carrying out correlation detection, on the other hand, also directly increase application cost, limit market application and the popularization of this technology.
Summary of the invention
The object of the invention is to overcome prior art deficiency, provide a kind of quick and precisely, the TORCH of visual result detects gene chip and test kit.
For solving the problems of the technologies described above, technical scheme of the present invention is:
One group of probe being used for TORCH and detecting, comprises toxoplasma gondii TOX probe, rubella RV probe, cytomegalovirus HCMV probe, cytomegalovirus HCMV probe and hsv HSV II type probe; Described probe sequence is:
Toxoplasma gondii TOX probe: SEQ ID NO.1;
Rubella RV probe: SEQ ID NO.2;
Cytomegalovirus CMV probe: SEQ ID NO.3;
Hsv HSV I type probe: SEQ ID NO.4;
Hsv HSV II type probe: SEQ ID NO.5.
Further, content of the present invention also comprises a kind of gene chip detected for TORCH, comprises solid-phase and above-mentioned probe.
Further, content of the present invention also comprises a kind of gene chip kit detected for TORCH, comprises above-mentioned gene chip.
Further, the gene chip kit that described TORCH detects, also comprise following primer pair, toxoplasma gondii TOX primer pair, rubella RV primer pair, cytomegalovirus CMV primer pair, hsv HSV primer pair, the sequence of described primer pair is respectively:
Toxoplasma gondii TOX primer pair SEQ ID NO.6 and SEQ ID NO.7;
Rubella RV primer pair SEQ ID NO.8 and SEQ ID NO.9;
Cytomegalovirus CMV primer pair SEQ ID NO.10 and SEQ ID NO.11;
Hsv HSV primer pair SEQ ID NO.12 and SEQ ID NO.13.
Further, on described primer containing there being biotin labeling.
Further, the gene chip kit that described TORCH detects also comprises PCR reaction system, negative controls, positive reference substance, hybridization buffer, washings, BW reaction solution and TMB nitrite ion.
Further, described negative controls is physiological saline, described positive reference substance is the mixed solution of TOXO plasmid, RV plasmid, CMV plasmid, HSV I plasmid and HSV II plasmid containing goal gene, and described goal gene is respectively as shown in sequence table SEQ ID NO.14 ~ 18.
Further, described hybridization buffer is trisodium citrate 0.3M, sodium-chlor 3M, sodium lauryl sulphate 0.3M, Triton X – 1000.1% aqueous solution.
Further, described washings is divided into washings A and washings B, and described washings A is 0.01-0.1N NaOH solution, and described washings B is 0.1 × SSC solution.
Further, described BW reaction solution is be diluted to suitable concentration with the affinity element that HRP marks by 20 × SSC damping fluid.
The present invention, by the selection of specific primer and probe, in conjunction with biochip technology, not only increases detection efficiency and accuracy, meets the demand of the Site Detection of hospital, antenatal detection etc.
The present invention is increased to pathogenic agent DNA by round pcr, and the oligonucleotide probe on amplified production and biochip is hybridized, and by by biotin labeling, after nitrite ion develops the color, shows detected result intuitively.Gene chip kit of the present invention, is amplified by the visible light optical of biochip, has accomplished that signal resolution is high, and signal can be taken pictures with ordinary digital camera or the direct interpretation of naked eyes.Relative conventional fluorescent mark has good effect.
In order to understand better and implement, describe the present invention in detail below in conjunction with accompanying drawing.
Accompanying drawing explanation
Accompanying drawing 1-5 is for utilizing pathogenic agent plasmid amplification results of hybridization in genechip detection Torch 5 of the present invention.Wherein Figure 1A, B, C, D are respectively TOX plasmid concentration is 2 × 10
-1, 2 × 10
-2, 2 × 10
-3, 2 × 10
-4time, namely in system, template content is 4 × 10
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7detected result during ng, E is negative control group.It is 4 × 10 that Fig. 2 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of RV plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 3 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of CMV plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 4 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of HSVI plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 5 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of HSV II plasmid during ng, E is negative control group.This detection method, detection sensitivity is high, can detect the template of 2 × 10-6ng/ul, namely detects and is limited to 2 × 10
-4ng/ml.
Accompanying drawing 6-10 is the result utilizing genechip detection many groups clinical sample of the present invention.Wherein, Fig. 6 is extraction 6 groups of TOX infected specimen DNA, obtains result by carrying out hybridization after pcr amplification; Fig. 7 is extraction 6 groups of RV infected specimen DNA, obtains result by carrying out hybridization after pcr amplification; Fig. 8 is extraction 6 groups of cmv infection samples, obtains result by carrying out hybridization after pcr amplification; Fig. 9 is extraction 6 groups of HSVI infected specimens, obtains result by carrying out hybridization after pcr amplification; Figure 10 is extraction 6 groups of HSV II infected specimens, obtains result by carrying out hybridization after pcr amplification.
Figure 11 increases after five kinds of pathogenic agent are extracted DNA mixing, and utilizes gene chip to detect, the result obtained.
Embodiment
The determination of [embodiment 1] target fragment, primer and probe
According to the gene order of the toxoplasma gondii (Toxopasma) of report, rubella virus (RubellaVirus), cytomegalovirus (Cytomegalo Virus), herpes simplex virus I and II type (Herpes VirusI/II), select one section of specificity is better, fragment length is applicable to gene order as the target sequence of detection;
The target sequence of TOX is chosen as the B1 Gene Partial sequence (SEQ ID NO.14) of toxoplasma gondii high conservative, research shows, can not detect B1 gene in people and relative species, and in 21 Toxoplasma gondii Strains, all can detect B1 gene, B1 gene has high degree of specificity.
The target sequence of RV virus is chosen as the gene (SEQ ID NO.15) of encoded packets membrane glycoprotein E1, and E1 gene has well-conserved relative to other fragments.
The target sequence of CMV is chosen as the UL123 gene (SEQ ID NO.16) of coding IE1.The IE1 of this genes encoding to virus copy and pathogenecity plays a part very important.
The target sequence of HSVI and HSVII is chosen as the UL30 gene order conservative region of encoding hsv dna polymerase catalytic subunit, is respectively (SEQ ID NO.17) and (SEQ ID NO.18).UL30 gene is the DNA polymerase gene of hsv, copies virus genomic and play an important role in virus multiplication.
According to target sequence, after target sequence is determined, according to primer and probe design principle, each pathogenic agent primer and the concrete sequence of probe of design are as follows, carry out to make chip results after nitrite ion develops the color, directly by the interpretation of naked eyes, to mark at R primer 5 ' end vitamin H:
Tox
F primer: 5 '-TTTGCATAGGTTGCAGTCACT (SEQ ID NO.6)
R primer: 5 '-biotin-GACCAATCTGCGAATACACCAA (SEQ ID NO.7)
TOX probe: AAAAAAAAAAAAAAAAAAAAATTCATGAGTATCTGTGCAAC (SEQ ID NO.1)
RV：
F primer: 5 '-GAACACCCGTTCTGCAACACG (SEQ ID NO.8)
R primer: 5 '-biotin-ATGGCGTTGGCAAACCGGGGAG (SEQ ID NO.9)
RV probe: AAAAAAAAAAAAAAAAAAAAATCGGGAGCCCGAATTGCCACGGC (SEQ ID NO.2)
CMV:
F primer: 5 '-GGAGAGCAGACTCTCAGAGGA (SEQ ID NO.10)
R primer: 5 '-biotin-CGCCGCATTGAGGAGATCTC (SEQ ID NO.11)
CMV probe: AAAAAAAAAAAAAAAAAAAAAAGACCTTCATGCGACGAGATCTCCTCAAT (SEQ ID NO.3)
HSV：
F primer: 5 '-AGGCGCCCAAGCGTCCGG (SEQ ID NO.12)
R primer: 5 '-biotin-GCTGGGGTACAGGCTGGCA (SEQ ID NO.13)
HSV1 probe: AAAAAAAAAAAAAAAAAAAAAGGGCGGGGGCGAGCGGGAGCCGGA (SEQ ID NO.4)
HSV2 probe: AAAAAAAAAAAAAAAAAAAACGAGGACGGGGACGAGGACGGGGACG (SEQ ID NO.5)
The preparation of [embodiment 2] chip
By vacuum gas-phase precipitator method plated film, rotatory vacuum coating equipment is used to be about 2.5mm at thickness, diameter 20cm silicon chip (SiO2) plates the film of the silicon nitride of 475 dusts and the TSPS of 135 dusts, prepare corresponding biosensor, and cover the Poly(Phe)-Methionin coating of 150 dusts, eventually pass 1-10uM hydrochloric acid bromosuccinimide base nicotinate process 20 minutes, clear water is cleaned for chip manufacturing.
By amido modified probe, arrange on the chip managed everywhere by probe point sample respectively according to table 1, often group establishes two parallel point of samples: react under room temperature, prepares the gene chip comprising probe.
Table 1TORCH chip matrix
Contrast PolyA probe sequence is 5 '-biotin-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
On the chip amido modified PolyA probe point sample managed everywhere according to chip matrix, react under room temperature, prepare the gene chip comprising probe.
[composition of embodiment 3 test kit and detection method]
Test kit forms:
1, extract: test kit composition of the present invention does not comprise DNA/RNA and extracts reagent, and agents useful for same is that the paramagnetic particle method viral DNA/RNA of TIANGEN Biotech (Beijing) Co., Ltd. extracts test kit, and article No. is DP438.
2, increase: PCR system, component is in table 2, table 3
Wherein shown in CMV, HSV, TOX system table 2 composed as follows:
Table 2TOX, CMV, HSVI, HSV II amplification system composition
| Name of material | Concentration | Add-on |
| 5×Buffer | 5× | 5ul |
| dUTPmix | 25μM | 0.2ul |
| F-primer | 200μM | 0.025 |
| R-primer | 200μM | 0.025 |
| Taq enzyme | 5U/ul | 0.25ul |
| UNG enzyme | 1U/ul | 0.2ul |
| ddH2O | Add water and mend 23ul |
(for TOX detection system F-primer in table: TOX-F primer, TOX-R-primer.CMV detection system F-primer: CMV-F primer, CMV-R-primer.HSV detection system F-primer: HSV-F primer, HSV-R-primer.)
RV is retrovirus, and system proportioning is as shown in table 3 below:
Table 3RV rubella virus amplification system
Add UNG enzyme in reaction system and can remove amplified production pollution, reduce false positive results; Taq enzyme, dUTP mixed solution, post transcription cloning reagent, UNG enzyme are purchased from Shenzhen Fei Peng Biological Co., Ltd..
3, hybridize:
Material:
Negative controls: detecting TORCH is negative physiological saline); Positive control: the mixed solution of TORCH gene C MV plasmid, HSV I plasmid, HSV II plasmid, TOXO plasmid, RV plasmid, is specially the plasmid (Nanjing Genscript Biotechnology Co., Ltd.'s synthesis) will built after goal gene segment condense; Hybridization buffer: trisodium citrate 0.3M, sodium-chlor 3M, sodium lauryl sulphate 0.3M, Triton X – 1000.1% aqueous solution liquid); Washings A (0.01-0.1N NaOH); Washings B (0.1 × SSC); BW reaction solution: the affinity element marked by HRP with 20 × SSC damping fluid is diluted to suitable concentration; Wherein 20 × SSC:(NaCl:3M, trisodium citrate: 0.3M); TMB nitrite ion is (the W-TMB colouring reagents that Sangon Biotech (Shanghai) Co., Ltd. buys, article No.: C510025-0005); According to the gene chip that embodiment 2 is obtained
Detection method:
1. PCR process: 50 DEG C of 2min; 94 DEG C of 5min; 94 DEG C, 30s--56 DEG C, 30s--72 DEG C, 30s, carry out 40 circulations; 72 DEG C, for subsequent use after 10min
2. by pcr amplification product 95 DEG C heating 3 minutes, newly, be placed in rapidly on ice;
3. get on 10ul pcr amplification product to prepared biochip;
4. 100ul hybridization buffer is got on chip, 60 DEG C of reactions 60 minutes;
5. 3 times are washed, each 5-10s by the washings solution A of preheating;
6. chip is placed in 50 DEG C of washings B and washs 1min, air blow drying chip surface;
7. BW reaction solution 100ul is got on chip, room temperature reaction 10 minutes;
8. 3 times are cleaned by washings B solution, each 1 minute, air blow drying chip surface;
9. get 100ul TMB nitrite ion in chip surface, react 5 minutes;
10. clean 3 times by washings B solution, each 1 minute, air blow drying chip surface, photographic camera is taken pictures or naked eyes are read
Read signal.
[embodiment 4 sensitivity analysis]
1. method: utilize the plasmid HSVI plasmid containing target fragment, HSV II plasmid, CMV plasmid, TOXO plasmid, RV plasmid (Nanjing Genscript Biotechnology Co., Ltd.'s synthesis) are tested to gene chip prepared by embodiment 2: synthetic plasmid is 2ug, by plasmid 1ml TE buffer solution, plasmid concentration is 2ug/ml and 2000ng/ml, and various high density plasmid is carried out gradient dilution according to 10 times, and it is as shown in table 4 below that dilution obtains plasmid concentration:
Table 4 dilutes group and the concentration of rear plasmid to be detected
Different plasmid is added corresponding system increase, system cumulative volume is 25ul, and adding plasmid template amount is 2ul.
According to the testing process of embodiment 3, utilize gene chip of the present invention, the amplification of the plasmid of above-mentioned different concns is detected.
2. result: Fig. 1-5 is for utilizing genechip detection Torch5 kind pathogenic agent plasmid amplification results of hybridization of the present invention.Wherein Figure 1A, B, C, D are respectively TOX plasmid concentration is 2 × 10
-1, 2 × 10
-2, 2 × 10
-3, 2 × 10
-4time, namely in system, template content is 4 × 10
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7detected result during ng, E is negative control group.It is 4 × 10 that Fig. 2 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of RV plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 3 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of CMV plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 4 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of HSVI plasmid during ng, E is negative control group.It is 4 × 10 that Fig. 5 A, B, C, D are respectively template content in system
-4ng, 4 × 10
-5ng, 4 × 10
-6ng and × 10
-7the detected result of HSV II plasmid during ng, E is negative control group.This detection method, detection sensitivity is high, can detect the template of 2 × 10-6ng/ul, namely detects and is limited to 2 × 10
-4ng/ml.
[analysis of embodiment 5 recall rate]
1. sample: get 6 groups of clinical infections TOX, RV, CMV, HSVI, HSV II sample respectively, extract DNA, after pcr amplification, utilize gene chip of the present invention to detect.
2. result: as Fig. 6-10, utilize gene chip provided by the invention can detect whole 36 parts of positive sample, recall rate is 100%.Wherein, Fig. 6 is extraction 6 groups of TOX infected specimen DNA, obtains result by carrying out hybridization after pcr amplification; Fig. 7 is extraction 6 groups of RV infected specimen DNA, obtains result by carrying out hybridization after pcr amplification; Fig. 8 is extraction 6 groups of cmv infection samples, obtains result by carrying out hybridization after pcr amplification; Fig. 9 is extraction 6 groups of HSVI infected specimens, obtains result by carrying out hybridization after pcr amplification; Figure 10 is extraction 6 groups of HSV II infected specimens, obtains result by carrying out hybridization after pcr amplification.
3. hybrid detection: increase after five kinds of pathogenic agent being extracted DNA mixing, and utilize gene chip to detect, obtain result as 11 figure, from the results of view, except above independent cause of disease physical efficiency accurately being detected, under 5 kinds of pathogenic agent mixing exist situation, also can accurately detect.
In sum, the present invention is increased to pathogenic agent DNA by round pcr, and the oligonucleotide probe on amplified production and biochip is hybridized, and by by biotin labeling, after nitrite ion develops the color, shows detected result intuitively.Gene chip kit of the present invention, is amplified by the visible light optical of biochip, has accomplished that signal resolution is high, and signal can be taken pictures with ordinary digital camera or the direct interpretation of naked eyes.Relative conventional fluorescent mark has good effect.The present invention, by the selection of specific primer and probe, in conjunction with biochip technology, not only increases detection efficiency and accuracy, meets the demand of the Site Detection of hospital, antenatal detection etc.
It should be noted that, above-mentionedly only to describe the present invention with preferred embodiment, interest field of the present invention can not be limited at this point, therefore when not departing from inventive concept, the equivalence that the content of all utilizations specification sheets of the present invention and accompanying drawing part is carried out changes, and all should be included in right of the present invention.
Claims (10)
1. one group of probe being used for TORCH and detecting, is characterized in that: comprise toxoplasma gondii TOX probe, rubella RV probe, cytomegalovirus HCMV probe, cytomegalovirus HCMV probe and hsv HSV II type probe; Described probe sequence is:
Toxoplasma gondii TOX probe: SEQ ID NO.1;
Rubella virus RV probe: SEQ ID NO.2;
Cytomegalovirus CMV probe: SEQ ID NO.3;
Hsv HSV I type probe: SEQ ID NO.4;
Hsv HSV II type probe: SEQ ID NO.5.
2., for the gene chip that TORCH detects, it is characterized in that: comprise probe groups described in solid-phase and claim 1.
3., for the gene chip kit that TORCH detects, it is characterized in that: comprise gene chip according to claim 2.
4. the gene chip kit of TORCH detection according to claim 3, it is characterized in that: also comprise following primer pair, toxoplasma gondii TOX primer pair, rubella RV primer pair, cytomegalovirus CMV primer pair, hsv HSV primer pair, the sequence of described primer pair is respectively:
Toxoplasma gondii TOX primer pair SEQ ID NO.6 and SEQ ID NO.7;
Rubella RV primer pair SEQ ID NO.8 and SEQ ID NO.9;
Cytomegalovirus CMV primer pair SEQ ID NO.10 and SEQ ID NO.11;
Hsv HSV primer pair SEQ ID NO.12 and SEQ ID NO.13.
5. the gene chip kit of TORCH detection according to claim 4, is characterized in that: containing there being biotin labeling on described primer.
6. the gene chip kit that the TORCH according to claim 4 or 5 detects, is characterized in that: also comprise PCR reaction system, negative controls, positive reference substance, hybridization buffer, washings, BW reaction solution and TMB nitrite ion.
7. the gene chip kit of TORCH detection according to claim 6, it is characterized in that: described negative controls is physiological saline, described positive reference substance is the mixed solution of TOXO plasmid, RV plasmid, CMV plasmid, HSV I plasmid and HSV II plasmid containing goal gene, and described goal gene is respectively as shown in sequence table SEQ ID NO.14 ~ 18.
8. the gene chip kit of TORCH detection according to claim 6, is characterized in that: described hybridization buffer is trisodium citrate 0.3M, sodium-chlor 3M, sodium lauryl sulphate 0.3M, Triton X – 1000.1% aqueous solution.
9. the gene chip kit of TORCH detection according to claim 6, it is characterized in that: described washings is divided into washings A and washings B, described washings A is 0.01-0.1N NaOH solution, and described washings B is 0.1 × SSC solution.
10. the gene chip kit of TORCH detection according to claim 6, is characterized in that: described BW reaction solution is be diluted to suitable concentration with the affinity element that HRP marks by 20 × SSC damping fluid.
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