A kit for rapid detection of il28b SNP rs12979860 polymorphism
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A kit and polymorphic technology, applied in the biological field, achieve the effect of short detection process, high degree of automation, and easy operation
Active Publication Date: 2011-12-14
李艳 +1
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[0008] At present, there is no literature report on the kit for detecting IL28B SNP rs12979860 by common PCR reaction
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Embodiment 1
[0038] The preparation of embodiment 1 kit of the present invention
[0039] (1) The composition of peripheral blood genomic DNA extraction reagents is as follows: deionized water, 6M NaI, chloroform / isoamyl alcohol (24:1, v / v) mixture, isopropanol and absolute ethanol.
[0040] (2) Primers:
[0041] TT genotype upstream amplification primer sequence is:
[0042] 5'-TCGCCAGGGCCCCTAACCTC-3' (SEQ ID No.1)
[0043] TT genotype downstream amplification primer sequence is:
[0044] 5'-GGAGTGCAATTCAACCCTGGTTCA-3' (SEQ ID No.2)
[0045] CC genotype upstream amplification primer sequence is:
[0046] 5'-GAGGATCCCTCCTGGGGCGG-3' (SEQ ID No.3)
[0047] The downstream amplification primer sequence of CC genotype is:
[0048] 5'-GGAGTGCAATTCAACCCTGGTTCG-3' (SEQ ID No.4)
[0070] Preliminary experiments show that there is no difference between the sensitivity, specificity and sensitivity of this kit compared with the sequencing method, which fully meets the practical requirements of current clinical diagnosis and treatment:
[0071] Table 1 Comparison of two methods for detecting rs12979860 polymorphism
[0072]
[0073]
[0074] in:
[0075] ① Specificity: 100%;
[0076] ②Sensitivity: 100%;
[0077] ③ Positive predictive value: the positive predictive value reaches 100%;
[0078] ④ Negative predictive value: the negative predictive value reaches 100%;
[0079] ⑤Repeatability: The results of repeated experiments are consistent;
[0080] ⑥ Time-consuming: The detection time of a clinical specimen is about 2 to 3 hours, which is short.
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Abstract
The invention relates to a kit for rapidly detecting single nucleotide polymorphism (SNP) rs12979860 of an IL28B gene. By the kit, a specific primer is designed, the specific primer and template deoxyribonucleic acid (DNA) in a sample are subjected to an ordinary polymerase chain reaction (PCR), and the polymorphic forms (CC, CT and TT) of IL28B SNP rs12979860 of the sample is judged according to the result of the PCR, so that a result of therapeutic effect prediction is provided for the clinical antiviral therapy of hepatitis C patients. The kit is easy to operate, can detect the polymorphism quickly, is economic and effective, can be widely used for screening the therapeutic effect of the hepatitis C patients before antiviral therapy, is convenient for clinicians to formulate therapeutic schedules pertinently, has high clinical application value, and is particularly suitable for clinical promotion.
Description
technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for rapidly detecting IL28B SNP rs12979860 polymorphism, which is used for predicting the antiviral treatment effect of hepatitis C patients. Background technique [0002] Hepatitis C infection is an infectious disease that seriously endangers human health, and its prevalence is distributed worldwide. According to WHO statistics, there are about 140 million to 170 million hepatitis C virus (Hepatitis C virus, HCV) infected people in the world. The infection rate of HCV in my country is about 3.2%, about 38 million HCV infected people, and about 35,000 new cases of hepatitis C every year. [0003] Previous studies have shown that the treatment method and therapeutic effect of hepatitis C are closely related to the virus genotype. Patients with genotype 1 and genotype 4 are treated with pegylated interferon and ribavirin (PegIFN / RBV)48 weeks; genotype 2 and genotype 3 patients we...
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