Displacement amplification method for detecting hepatitis B viruse P gene YMDD variation

Abstract

The invention relates to a displacement amplification process which is used to detect hepatitis b virus P gene YMDD variation, which is characterized in that detected HBV P gene YMDD motif and variation YIDD, wherein YVDD sequence is displaced into indication DNA through hybridization-ligase reaction, and then is reversely amplified into an induction system. The YMDD hybrid sequence is separated into left half and right half and is added on the front end and the tail end of a section of induction DNA, is hybridized with P gene YMDD region sequence after added with front and tail induction DNA, make the front end and tail end are in accordance with each other, and the hybrid gap is connected with ligase. The reverse amplification induction DNA adopts right middle sequence of induction DNA as primer, reversely amplifies the covalence connection of the front end and the tail end to induct DNA, wherein base groups on the two sides of the gap are connected through matching with dependency, thereby being suitable to one nucleotide mutagenic gene analysis. Detection can be displaced into several inducted PCR, thereby avoiding the recontamination of one set of system amplification products. Detected RNA can be displaced into induction DNA to be directly detected through hybridization-connection reaction, or is suitable to the real-time quantitative analysis of detected gene through combining with SYBR Green dye or fluorescent molecular probe.

Description

Technical field: [0001] The invention relates to a molecular detection method of the hepatitis B virus (Hepatitis B Virus HBV) gene and its variation. Especially HBV genome includes DNA and RNA and YMDD gene mutation polymerase chain reaction (Polymerase Chain Reaction) PCR amplification detection method and detection kit. Background technique: [0002] Hepatitis B is an infectious disease that seriously threatens human health. According to the report of the World Health Organization, about 2 billion people in the world have been infected with HBV, of which more than 350 million people are chronic hepatitis B virus HBV infected people. my country is a high prevalence area of ​​hepatitis B infection . Hepatitis B virus belongs to the hepadnaviridae family and is also the main cause of chronic hepatitis, cirrhosis, liver failure and primary hepatocellular carcinoma. The diagnosis of hepatitis B infection is currently mainly based on serological enzyme immunoassay detection. W...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies in the above fields, a displacement amplification method for detecting the variation of the hepatitis B virus P gene YMDD is provided, and the amplification sensitivity is reduced by the displacement method to avoid the false positive problem caused by the recontamination of the PCR product, and at the same time, the fluorescent color is developed , can detect the content of each mutant strain

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