Reagent kit for multi-channel fluorescent PCR detection of polymorphism of IL28B gene loci

Abstract

The invention discloses a reagent kit for multi-channel fluorescent PCR detection of the polymorphism of IL28B gene loci. The reagent kit can specifically detect the polymorphism of the IL28B gene loci rs12979860 and rs8099917, the two loci are simultaneously detected in one PCR reaction pipe, and the specific genotype is made clear and definite. Besides, operation is easy, convenient and fast, the result is objective and high in controllability, efficiency is high, cost is low, contamination is avoided through closed-pipe operation, and the reagent kit is suitable for a multi-channel fluorescent PCR instrument and is a powerful product for clinically detecting the polymorphism of the IL28B gene loci.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a kit for detecting the polymorphism of the IL28B gene locus based on a multi-channel fluorescent PCR method. Background technique [0002] Hepatitis C is a widespread infectious disease worldwide. According to statistics, there are currently about 180 million people in the world who have been infected with HCV virus, accounting for about 3% of the total population. Hepatitis C is mainly transmitted by blood, and acupuncture and drug abuse are also common routes of infection. Many studies have shown that the prognosis of hepatitis C and the efficacy of antiviral therapy are closely related to the polymorphism of the host IL28B gene. [0003] The IL28B gene is located on chromosome 19 and consists of 6 exons and 5 introns. Its encoded product IFN-λ3 belongs to the type III interferon family and participates in the response to antiviral therapy, thus affecting the spontaneous clearan...

AI Technical Summary

Problems solved by technology

The melting curve method has high requirements on the temperature control of the instrument, and requires the instrument to have the function of high-resolution melting curve analysis (HRM), but our commonly used fluorescent quantitative PCR instruments generally lack this function; the ARMS-PCR method requires electrophoresis of PCR products Analysis, not suitable for clinical application; the sequencing method has high accuracy, but the instrument is expensive, the operation is complicated and time-consuming, and has high requirements for operators and experimental sites

[0006] The Chinese patent "IL28B gene polymorphism detection kit based on real-time fluorescent PCR" application number is 201510007333.1 discloses a real-time fluorescent PCR-based IL28B gene polymorphism detection kit, two polymorphic sites each have a reaction tube Liquid, that is, each sample requires two tubes of reaction solution detection, the first disadvantage is that each sample needs to occupy two reaction wells, which halves the throughput

The second disadvantage is that it is inconvenient to operate. If one of the sites is detected first, the other site needs to be tested strictly in accordance with the original order of the sample. When the sample is detected at two sites at the same time, the order of adding different reaction solutions must be strictly recorded when adding the reaction solution to avoid misjudgment of results

In addition, it can be seen from the drawings published in its patent that there is obvious non-specific amplification in the amplification curve of its homozygote, which can be easily interpreted as a heterozygote, which is related to the design of the primer probe and the probe luminescent group and The selection of the quenching group has a great relationship, and it is difficult for operators without interpretation experience to accurately interpret the results

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