Method for identifying HBV gene mutation type, special chip and reagent kit

Abstract

The invention discloses a method for identifying HBV gene mutation type and a chip and a reagent box which are especially used with the method. The method for identifying the HBV gene mutation type takes the detected the genome of the hepatitis B virus as the template and uses the following primer group and a DNA polymerase which has no exonuclease activity from the 3'end to 5' end for implementing multiple PCR magnification; the primer group comprises a universal primer and a or more than one special primers; the special primer 5' end is a bar code sequence and the length of the bar code sequence is 5 to 25 nt; the bar code sequence has a comparatively low homology with the hepatitis B virus; the special primer totally matches with the corresponding segment of the hepatitis B virus comprising wild base group or mutant base group at a mutation point; the gene chip comprises a plurality of probes and the probes are corresponding to the bar code sequence one to one and every probe only contains one bar code sequence.

Description

technical field [0001] The invention relates to a method for identifying HBV gene mutation types in the field of gene analysis, as well as a dedicated chip and a kit. Background technique [0002] Chronic hepatitis B is one of the most serious health problems at present. About 2 billion people in the world have been infected with hepatitis B virus (HBV), of which 350 million people are chronic HBV infected people, and about 1 million people die every year. Liver failure, liver cirrhosis and primary hepatocellular carcinoma (HCC) caused by HBV infection. my country is a high prevalence area of ​​HBV infection. There are 130 million hepatitis B virus carriers and more than 30 million hepatitis B patients in China. The positive rate of HBsAg in the general population was 9.09%. HBV is a highly mutated virus. During its reverse transcription and replication process, due to the lack of correction function of RNA polymerase and reverse transcriptase, the virus can undergo one or...

AI Technical Summary

Problems solved by technology

However, the reagents are expensive, and there are a small number of problems that are not easy to interpret the results due to DNA sequence polymorphisms, so they are rarely used in clinical testing; (7) Sequence Specific Oligonucleotide Probe (SSOP) gene chip Technology: Utilize the high-throughput and parallel detection characteristics of the gene chip to design probes for many genes that cause mutations. These probes are combined on the same chip. Needle hybridization, according to the hybridization signal to determine the mutant genotype of HBV

This technology is difficult to accurately distinguish single base differences, and the specificity is poor; at the same time, it is difficult to accurately distinguish mutant / wild-type co-existing individuals

[0008] The above-mentioned technologies all have a common deficiency, that is, any single method cannot detect HBV gene mutation types in dominant populations / disadvantaged populations in a high-throughput, accurate and specific manner

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