PCR (Polymerase Chain Reaction) primer, PCR primer group, PCR detection probe and PCR detection kit for detecting hepatitis B virus as well as detection method

Abstract

The invention discloses a PCR (Polymerase Chain Reaction) primer, a PCR primer group, a PCR detection probe and a PCR detection kit for detecting hepatitis B virus as well as a detection method. The kit contains the high-sensitivity detection primer group and the high-sensitivity detection probe and can be used for detecting single copy HBVDNA (Hepatitis B virus-Deoxyribonucleic acid), the limit of detection of the kit in a serum specimen can reach 2IU / mL, and compared with an imported reagent Roche COBAS, the kit is relatively sensitive. The detection method disclosed by the invention can be used for detecting different subtypes of hepatitis B pathogens around the world; due to addition of an internal control system in the detection method, false negative can be effectively prevented; due to addition of a UNG (uracil-N-glycosylase) system in the detection method, pollution can be effectively avoided; the kit and the detection method have the beneficial effects that the kit and the detection method can be applied to diagnosis of hepatitis B, evaluation of research, development and screening of anti-hepatitis-B new drugs and evaluation of an anti-hepatitis-B treatment effect and relapsing and have wide clinical application effects, and the popularization and application of the kit and the detection method are facilitated.The invention discloses a PCR (Polymerase Chain Reaction) primer, a PCR primer group, a PCR detection probe and a PCR detection kit for detecting hepatitis B virus as well as a detection method. The kit contains the high-sensitivity detection primer group and the high-sensitivity detection probe and can be used for detecting single copy HBV DNA (Hepatitis B virus-Deoxyribonucleic acid), the limit of detection of the kit in a serum specimen can reach 2IU / mL, and compared with an imported reagent Roche COBAS, the kit is relatively sensitive. The detection method disclosed by the invention can be used for detecting different subtypes of hepatitis B pathogens around the world; due to addition of an internal control system in the detection method, false negative can be effectively prevented; due to addition of a UNG (uracil-N-glycosylase) system in the detection method, pollution can be effectively avoided; the kit and the detection method have the beneficial effects that the kit and the detection method can be applied to diagnosis of hepatitis B, evaluation of research, development and screening of anti-hepatitis-B new drugs and evaluation of an anti-hepatitis-B treatment effect and relapsing and have wide clinical application effects, and the popularization and application of the kit and the detection method are facilitated.

Description

technical field [0001] The invention belongs to the field of virus detection, and relates to a PCR primer, a primer set, a probe, a kit thereof and a detection method for detecting hepatitis B virus. Background technique [0002] Hepatitis B virus (HBV) infection is distributed globally, with more than two billion infected people, and it is one of the most serious infectious diseases in my country. According to the national epidemiological survey of hepatitis B carried out in 2010, there are still about 93 million hepatitis B surface antigen carriers in my country, and hepatitis B and liver cancer patients account for about a quarter of the world. Chronic HBV infection is the main cause of chronic hepatitis, liver cirrhosis, liver failure and primary liver cancer, and causes more than 1 million deaths every year, causing great social and economic harm. [0003] HBV replication is the root cause of chronic HBV activity. Effective antiviral therapy can delay the progression o...

AI Technical Summary

Problems solved by technology

[0005] At present, most hospitals in China generally use domestic fluorescent quantitative PCR reagents to detect serum HBV DNA load, which are cheap, but the sensitivity is poor, and the lower limit of detection is generally 100 IU / ml, for HBV DNA loads below this lower limit, no further distinction can be made, and for higher HBV DNA loads, the detection accuracy is poor, and the detection values ​​are often lower than the actual level

At present, the imported reagents used in China mainly include Roche’s COBAS amplicor and COBAS Taqman, among which COBAS Taqman HBV DNA is the only automatic control technology for test tube diagnosis using Taqman technology approved by the US FDA, and is included in the clinical practice guidelines for chronic hepatitis B. The recommended HBV DNA detection method has the advantages of high sensitivity, wide linear range, real-time monitoring, and good repeatability. The detection range can reach 20 IU / mL~1.7×108 IU / mL. Its equipment is expensive and the detection cost is high, so it is difficult to be widely used in clinical practice.

Moreover, HBV DNA levels at the time of discontinuation may be below the detection limit of Roche's reagent of 20 IU / mL

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