Preparation and application of guiding gene chip for HCV infection individual treatment

Abstract

The invention relates to a guiding gene chip for HCV infection individual treatment. The preparation method of the guiding gene chip comprises the following steps: preparing a universal primer; preparing an HCV hypotype nucleic acid parting probe and a human IL-28B rs12979860 investigation of polymorphism probe; preparing an oligonucleotide chip; building a polyfunctional RT-PCR system; building a hybrid system. The gene chip prepared by the method can discriminate five hypotypes of 1b, 2a, 3a, 3b and 6a of HCV at the same time, can detect polymorphism of CC, TT and CT three genes of human rs12979860, has the advantages that the gene chip is quick, accurate, and high in flux and specificity, and can provide guiding for HCV clinical diagnosis and individual treatment.

Description

Technical field [0001] The invention relates to the preparation and application of a guide gene chip for individualized treatment of hepatitis C virus infection, and belongs to the technical field of gene chip detection. Background technique [0002] Hepatitis C virus (hepatitis c virus, HCV) infection is a serious hazard. 170 million patients worldwide are infected with HCV, and there are 3 million new cases each year. 50-80% of people infected with HCV can develop a chronic state, of which 20-30% develop cirrhosis or liver cancer. There are differences in the geographical distribution of different genotypes or subtypes of chronic hepatitis C infection, the effect of antiviral therapy, and the severity of the disease. The genetic polymorphism of human IL-28B is closely related to the viral response rate of antiviral therapy. It is an important predictor of the success of treatment for hepatitis C patients. Individualized treatment of HCV infection can significantly increase th...

AI Technical Summary

Problems solved by technology

Serological methods are fast, simple, low-cost, and widely used clinically, but it is difficult to distinguish when there is mixed virus infection or virus mutation

[0005] 2) Direct sequencing method: direct sequencing using PCR products is the gold standard for typing, but the sensitivity is insufficient and limited by sequencing equipment, it is not easy to promote at the grassroots level

However, this method requires a special fluorescent quantitative PCR instrument and supporting reagents

[0007] 4) Nested PCR method: use two pairs of PCR primers to amplify the fragment, the first pair of PCR primers amplifies the fragment similar to ordinary PCR, the second pair of primers are nested primers, combined in the first PCR product, after two PCR program, enhanced amplification specificity, sensitivity and reliability are better than ordinary PCR, often used for low-concentration nucleic acid amplification, and its disadvantage is that it is easy to contaminate

This method has the advantages of sensitivity, high throughput, intensification, large-scale, and accurate results. However, due to its high technical requirements and the need for special detection instruments, there are still some restrictions in its promotion.

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