Detection kit of hepatitis B virus resistance to drug and hepatitis B virus genotype and detection method using the same

Abstract

The invention discloses a detection kit of hepatitis B virus resistance to a drug and a hepatitis B virus genotype and a detection method using the same. The kit is used based on a gene chip and contains three groups of primers. In detection, the three groups of primers respectively carry out multiple asymmetric PCR amplification processes on a sample to be detected, the three PCR amplification products are mixed, the mixture and a hybridization buffer solution are mixed to form a hybridization solution, the hybridization solution and a probe of the gene chip are subjected to hybridization and a mutant type and a genotype of the sample to be detected are determined through a fluorescence signal of the PCR product. The detection result shows that the kit has good specificity and high sensitivity and can simultaneously detect seven mutant types and three genotypes of samples to be detected. The seven mutant types comprise a 180M type, a 181T type, a 181V type, a 204I type, a 204V type, a 236T type and a 1896A type and the three genotypes comprise a HBV / B type, a HBV / C type and a HBV / D type.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit and detection method for drug resistance and genotype of hepatitis B virus. Background technique [0002] Hepatitis B virus is a hepatotropic DNA virus, which is one of the main pathogens causing acute and chronic hepatitis, and can lead to liver cirrhosis, liver failure and liver cancer. my country is a country with a high incidence of hepatitis B. In 2009, the bulletin of the Ministry of Health showed that the incidence of hepatitis B in 2008 was 1.0654 / 100,000, and the number of cases was 1.41 million. HBV infection is a threat to national health, and the early diagnosis and correct treatment of HBV should receive great attention. [0003] Interferon, lamivudine, telbivudine, adefovir and the soon-to-be-launched tenofovir are all first-line antiviral drugs in China. Strict correction mechanism leads to high nucleotide mismatch rate during HBV replication. Gene mu...

AI Technical Summary

Problems solved by technology

(1) Sequencing is the gold standard for HBV gene detection, with high accuracy, but low sensitivity, poor ability to detect mixed genotypes (need to screen a large number of clones), requires a sequencer and expensive reagents, and is not suitable for clinical detection; (2) RFLP method is not suitable for clinical detection. Complex equipment and reagents are required, but the standardization degree of the current method is poor; (3) the quantitative PCR method has high sensitivity, but the throughput is low, and it is difficult to detect the variation of multiple gene loci; Interpretation is easy to misjudge, the cost of detection is high (imported commercial kits), and it is difficult to carry out routinely in domestic clinical laboratories

The main obstacle to achieve this goal is that since the hybridization reaction between the labeled target nucleic acid and the oligonucleotide probe immobilized on the surface of the chip is the key factor affecting the detection of the chip, only one strand of double-stranded DNA can be combined with the immobilized oligonucleotide probe on the surface of the chip. The probe on the surface of the chip performs a hybridization reaction, and the other strand acts as an interference factor for the hybridization reaction due to competition

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