Multi-color fluorescent PCR detection kit for nucleic acid testing and genetic typing of hepatitis c virus and application

Abstract

The invention discloses a multi-color fluorescent PCR kit for nucleic acid testing and genetic typing of hepatitis c virus and an application thereof. The kit comprises RT-PCR reaction liquid, enzyme mixed liquid, HCV reaction liquid I, HCV reaction liquid II and HCV reaction liquid III, and the kit preferably also comprises HCV typing negative control, HCV typing positive control and application instruction. For the extracted HCV RNA, the hepatitis c virus RNA and the subtype of the hepatitis c virus in a test sample can be rapidly detected by adopting three-tube multi-color fluorescent PCR through one-step RT-PCR. The amplification method of the kit is simple, simplicity in operation is realized, the testing result is high in sensitivity, the specificity is good, the kit can be used for testing and genetic typing of HCV RNA in human serum or plasma, the judgment on the treatment difficulty can be favored, the establishment of an individual antivirus treatment scheme is favored, and the kit has the clinical popularization value.

Description

technical field [0001] The present invention relates to a disease pathogen gene detection kit, in particular to a kit for preparing hepatitis C virus and its genotyping detection using a multiple fluorescent PCR detection method, which is used for rapid and accurate detection of hepatitis C virus in clinical serum or plasma samples. Infection with hepatitis virus and the main domestic epidemic subtypes 1b, 2a, 3a, 3b, 6a. The invention belongs to the field of biotechnology. Background technique [0002] Hepatitis C virus (HCV) is the pathogen that causes hepatitis C and seriously endangers human health. It is mainly infected through blood transfusion, intravenous injection, mother-infant vertical, etc., and can cause chronic hepatitis, liver cirrhosis, and even liver cancer. According to the report of the World Health Organization, there are currently about 200 million people infected with HCV in the world (including at least 130 countries), accounting for about 3% of the ...

AI Technical Summary

Problems solved by technology

However, although the ELISA method used to detect HCV genotype-specific antigens can identify 6 genotypes, it cannot identify subtypes

And some studies suggest that serotyping may have limitations in specificity and sensitivity

The research methods of HCV RNA genotyping reported in domestic and foreign literatures in recent years mainly include HPLC enzyme digestion method, primer-specific electrophoresis typing method, melting point curve method, separate primer sequencing and other methods. Due to defects such as open-ended or multi-step management, it has not become a routine clinical technique

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