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Microdroplet-type digital PCR detection kit for latent hepatitis B virus

A detection kit and hepatitis B virus technology, applied in the biological field, can solve the problems of inaccurate and false negative OBI detection, and achieve the effects of efficient detection and accurate quantification, strong specificity and high detection efficiency

Pending Publication Date: 2019-05-21
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a dual-gene co-tube digital droplet PCR virus detection method and kit, which are used to solve the problem of low-load HBV infection and base mutation of HBV in the prior art, resulting in inaccurate detection of OBI , and even false negative results and other problems

Method used

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  • Microdroplet-type digital PCR detection kit for latent hepatitis B virus
  • Microdroplet-type digital PCR detection kit for latent hepatitis B virus
  • Microdroplet-type digital PCR detection kit for latent hepatitis B virus

Examples

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Comparison scheme
Effect test

Embodiment 1

[0119] Quantitative detection of HBV by dual-gene simultaneous digital PCR

[0120] 1. Dissolve the standard positive template in distilled water and dilute it to 6×10 2 copies / μL, 6×10 1 copies / μL, 6×10 0 copies / μL, 6×10 -1 copies / μL.

[0121] 2. Prepare a digital PCR reaction system (20 μL amplification reaction system): 10.0 μL digital PCR reaction solution, 0.5 μL (900 nM) forward primer of the S gene region primer set, 0.5 μL (900 nM) reverse primer of the S gene region primer set, Fluorescent probe 0.5μL (250nM), C gene region primer set forward primer 0.5μL (900nM), C gene region primer set reverse primer 0.5μL (900nM), fluorescent probe 0.5μL (250nM), primer probe set Respectively selected from combinations S1 and C1, 7.0 μL of templates; the template is one of the DNA of one of the above-mentioned samples to be tested, a negative reference product, and a positive reference product; the negative reference product is used as a negative control for the PCR reaction, ...

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Abstract

The invention provides a microdroplet-type digital PCR detection kit for a latent hepatitis B virus. The kit comprises digital PCR reaction liquid and a primer probe composition, and the primer probecomposition is selected from at least one of an S gene area primer probe composition and a C gene area primer probe composition. According to the kit, primer probes are respectively designed for S andC conserved sequences of the hepatitis B virus, a duplex double-gene digital PCR technology is adopted to synchronously detect HBV DNA in the same tube, and the operation is convenient and quick; thesensitivity and specificity of a detection method for the latent hepatitis B virus are improved, the probability that the false negative result is caused due to mutation is reduced as much as possible, and the detection efficiency can be further improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a dual-gene simultaneous tube droplet digital PCR virus detection method and a kit. Background technique [0002] HBV infection remains a major public health problem worldwide. Since hepatitis B virus infection was discovered, it has been paid more attention, especially in recent years, the potential clinical harm of hepatitis B virus infection has been paid more and more attention. The total number of people infected with HBV in the world was as high as 2 billion. Liver cirrhosis, primary liver cancer and the risk of liver failure, about 1 million people die each year from diseases related to HBV infection. In Asian countries such as China, India, South Korea, Singapore and Vietnam, 80-90% of liver cancer patients are caused by HBV infection. According to statistics, by 2030, about 10 million people in my country will die of chronic hepatitis B virus infection. HBV infection has b...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
Inventor 赵耀杨宇婷
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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