A hbv cccDNA digital PCR quantitative detection kit and its application

A quantitative detection and kit technology, applied in the field of virus in vitro nucleic acid detection, to achieve the effect of fast detection, high sensitivity, and ingenious detection design
CN104388598BActive Publication Date: 2016-08-24SHANGHAI CHROMYSKY MEDICAL RES
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHANGHAI CHROMYSKY MEDICAL RES
Publication Date
2016-08-24

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Abstract

The invention belongs to the technical field of virus in-vitro nucleic acid detection and particularly relates to a digital PCR quantitative detecting kit for HBV (hepatitis B virus) cccDNA and an application of the digital PCR quantitative detecting kit for HBV cccDNA. A digital PCR quantitative detecting probe composition of the HBV cccDNA is firstly designed, and the kit comprises a container for detecting the probe composition and a container for a purifying reagent containing liver puncture tissue DNA. Using of the kit comprise the following steps of extracting the DNA in the liver puncture tissue, and adopting PSAD (prostate specific antigen density) enzyme to digest the HBV cccDNA; by taking a digital PCR detecting chip as a reaction container and adopting a complex fluorescent multiple PCR primer, amplifying target DNA in detecting micro pores of the digital PCR chip; and obtaining the quantitative results of the HBV cccDNA in each cell according to a fluorescence signal in each micro pore of the digital PCR detecting chip. The digital PCR quantitative detecting kit for HBV cccDNA is strong in specificity, high in sensitivity, simple and convenient and quick. HBV cccDNA and HBV rcDNA are not needed to be separated and extracted, and the defects in the conventional HBV cccDNA detection are overcome, and therefore, the digital PCR quantitative detecting kit is suitable for large-scale popularization and application.
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Description

technical field

[0001] The invention belongs to the technical field of virus in vitro nucleic acid detection, and in particular relates to a HBV cccDNA digital PCR quantitative detection kit and an application thereof. Background technique

[0002] Chronic Hepatitis B (CHB) is a liver inflammatory disease caused by chronic infection of Hepatitis B Virus (HBV) in human liver cells, and its continuous progression can lead to liver cirrhosis and liver failure and liver cancer. It is estimated that there are currently about 350 million HBV carriers in the world, and about 1 million people die each year from liver failure, cirrhosis or liver cancer caused by HBV infection. In 2012, the "Guidelines for the Prevention and Treatment of Chronic Hepatitis B" jointly formulated by the Liver Disease Branch of the Chinese Medical Association and the Infectious Disease Branch of the Chinese Medical Association pointed out that there are currently about 93 million chronic HBV infections i...

Claims

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