Typing detecting kit and method for detecting digital PCR absolute quantification of HBV-B/C

A technology of HBV-B and detection kits, which is applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc. It can solve the problems of cross-hybridization reaction, low accuracy, complicated steps, etc., and achieve detection sensitivity and accuracy Accurate quantification, saving detection time, and simplifying the operation steps

Active Publication Date: 2016-03-30
ANHUI UNIV OF SCI & TECH
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Problems solved by technology

[0006] For the above-mentioned deficiencies that exist in the prior art, the technical problem to be solved in the present invention is: how to provide a kind of digital PCR absolute quantitative typing detection kit and detection method thereof for detecting HBV-B/C, for overcoming existing I

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  • Typing detecting kit and method for detecting digital PCR absolute quantification of HBV-B/C
  • Typing detecting kit and method for detecting digital PCR absolute quantification of HBV-B/C
  • Typing detecting kit and method for detecting digital PCR absolute quantification of HBV-B/C

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Embodiment Construction

[0040] In order to better understand the content of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the following specific examples are only used to illustrate the present invention, rather than limit the present invention.

[0041] 1. A digital PCR absolute quantitative typing detection kit for detecting HBV-B / C, comprising DNA extract, digital PCR reaction buffer A, digital PCR reaction buffer B, HBV-B viral gene positive quality control , HBV-C virus gene positive quality control, negative quality control and internal standard solution;

[0042] The DNA extract comprises 50mM Tris-HCl at pH 8.3, 0.5MNaCl, 5mM EDTA, CTAB with a mass concentration of 2.5% and DTT of 2mM;

[0043] The digital PCR reaction buffer A includes primers and probes of the HBV-B type and the primers and probes of the HBV-C type; the digital PCR reaction buffer B includes i...

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Abstract

The invention discloses a typing detecting kit and method for detecting the digital PCR absolute quantification of HBV-B/C. The kit comprises a DNA extraction solution, a digital PCR reaction buffer solution A, a digital PCR reaction buffer solution B, an HBV-B virus gene positive quality control, an HBV-C virus gene positive quality control, a negative quality control and an internal standard solution. The method includes the following steps of firstly, processing a to-be-detected sample; secondly, processing a quality control product; thirdly, preparing a digital PCR reaction mixed solution; fourthly, generating a micro-reaction liquid drop, and conducting digital PCR reaction amplification; fifthly, reading a fluorescent signal, analyzing an HBV type and calculating the number of copying times. By means of the digital PCR technology and the dual-color florescent probe, two subtypes of HBV are detected at the same time; without depending on an external standard object or a standard curve, the kit and method are easy and convenient to operate and can directly detect the HBV precise absolute quantification.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnostic biology, and in particular relates to a digital PCR absolute quantitative typing detection kit for detecting HBV-B / C and a detection method thereof. Background technique [0002] Hepatitis B virus is the pathogen that causes acute and chronic hepatitis B in humans. It can cause serious damage to liver cells, and persistent and stable HBV infection is closely related to the occurrence of liver cirrhosis and liver cancer, which is extremely harmful to human health. It is estimated that there are about 2 billion people who have been infected with HBV in the world, and more than 350 million infected people have persistent stable and chronic infections; the number of deaths from related diseases caused by HBV infection can reach 600,000-1 million every year. The prevalence of HBV infection varies widely around the world, with the highest prevalence in eastern Asia and low prevalence in n...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6851C12Q1/706C12Q1/707C12Q2600/166C12Q2563/159C12Q2545/113C12Q2563/107
Inventor 唐小龙程龙强刘新矿高毅吴汉霞蔡霞段舒婷王倩倩陈锐李运超金少鹏李辉屈爱军石磊铁牛
Owner ANHUI UNIV OF SCI & TECH
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