Kit for HCV virus genotyping detection, use method and application thereof

Abstract

The invention discloses a kit for HCV virus genotyping detection, a use method and an application thereof. The kit includes a nucleic acid amplification reagent and a hybridization reagent. The nucleic acid amplification reagent includes primers represented as the SEQ ID No. 1-4. The hybridization reagent includes probes represented as the SEQ ID No. 5-16. The use method of the kit includes the steps of: 1) nucleic acid extraction of a sample; 2) preparation of the amplification reagent; 3) PCR amplification; 4) a hybridization reaction; 5) a fluorescent reaction; 6) detection; and 7) determination on the genotype of HCV virus according to fluorescent signal value in the detection reaction system. The kit can be used for detecting nine different genotypes of HCV virus, has high flux, high sensitivity and high type coverage area, allows multi-target simultaneous detection and is simple in operation.

Description

technical field [0001] The invention relates to a genotyping detection kit and its use method and application, in particular to a hepatitis C virus genotyping detection kit and its use method and application. Background technique [0002] Hepatitis C virus (HCV), first discovered in 1989, is the most common blood transfusion and community-acquired non-A, non-B hepatitis virus in the world. HCV is a folded, single-stranded, positive-sense RNA virus that forms a separate genus within the Flaviviridae family. Worldwide, at least six genotypes and multiple subtypes and virus variants have been found. Infection with different genotypes can affect disease severity and response to treatment. HCV is mainly transmitted through contaminated blood and blood products, and the possibility of infection through human secretions is very low. [0003] The prevalence of HCV is over 3.0% worldwide, with a carrier rate of 3.7% in China. Most of the HCV in China are genotypes 1b and 2a, but ...

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Problems solved by technology

However, the liquid-phase chip method is slightly more complicated than the real-time fluorescent PCR operation, and needs to be opened after the PCR reaction is completed, which is a test for the anti-pollution ability of the product.

[0008] In addition, the operating cost of membrane hybridization method is relatively low, but the degree of automation is low, the time is long, and the sensitivity is not high

Real-time fluorescent PCR method can achieve high sensitivity, but it is difficult to realize the identification of multiple types due to the limitation of the number of channels

[0009] The above-mentioned detection methods all have their own defects, which bring certain difficulties to the diagnosis and treatment of clinical HCV types

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