Quantitative detection kit of hepatitis B virus (HBV) nucleic acid

A hepatitis B virus, detection kit technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as poor accuracy and low sensitivity, and achieve simple operation, low cost and good repeatability. Effect

Active Publication Date: 2014-03-19
东北制药集团辽宁生物医药有限公司
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Problems solved by technology

[0005] The purpose of the present invention is to provide a hepatitis B virus (HBV) nucleic acid quantitative detection kit, which fundamentally solves the problems of low sensitivity and poor accuracy in the existing hepatitis B virus (HBV) nucleic acid quantitative PCR technology, The TaqMan fluor

Method used

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  • Quantitative detection kit of hepatitis B virus (HBV) nucleic acid
  • Quantitative detection kit of hepatitis B virus (HBV) nucleic acid
  • Quantitative detection kit of hepatitis B virus (HBV) nucleic acid

Examples

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Embodiment 1

[0023] Real-time quantitative PCR detection example of HBV DNA

[0024] (1) Reagent preparation

[0025] a. Preparation of PCR reaction solution

[0026] The PCR buffer composition is 30mmol / L Tris-HCl (PH8.3), 10mmol / L (NH4) 2 SO 4 , 30mmol / LKCl, 4.5mmol / LMgCl 2 , 0.15mmol / L dNTPs. The concentration of the upstream primer and downstream primer for amplifying the target gene is 20 pmol, the concentration of the probe for target gene detection is 8 pmol, the concentration of the upstream primer and downstream primer of the internal standard is 8 pmol, and the concentration of the internal standard probe is 8 pmol.

[0027] b. Take the corresponding amount of Lysis Conjugate Solution and Internal Standard according to the proportion (400 μl of Lysis Conjugate Solution per person + 0.1 μl of internal standard per person), mix well to form Lysis Conjugate Solution-mix, centrifuge briefly and set aside.

[0028] c. According to the amount of the sample to be tested, negative c...

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Abstract

The invention discloses a quantitative detection kit of a hepatitis B virus (HBV) nucleic acid applied to the field of biomedical clinic diagnosis. The kit comprises a paramagnetic particle method extraction kit and an HBV nucleic acid amplification kit, wherein the paramagnetic particle method extraction kit comprises a pyrolysis binding solution, a rinsing solution, an eluant and magnetic bead liquid; the HBV nucleic acid amplification kit comprises an HBV-PCR (Polymerase Chain Reaction) reaction solution, an enzyme mixed solution, an HBV-interior label, HBV quantitative reference products 1-4, a negative quality product, a clinical positive quality product and a strong positive quality product. The quantitative detection kit is simple, convenient and fast in operation, low in cost, high in detection sensitivity, good in repeatability, high in conservative property of primer and probe, and strong in specificity, and covers different subtypes or variants of the hepatitis B virus, improvement of the accuracy and the specificity of the hepatitis B detection is facilitated, an efficient interior label system is led in, the problems such as reciprocal inhibition, interference and the like caused by simultaneous amplification of a target gene and the interior label are solved, the overall PCR amplification process can be effectively monitored, and a false negative result is avoided.

Description

technical field [0001] The invention relates to a hepatitis B virus (HBV) nucleic acid quantitative detection kit in the field of biomedical clinical diagnosis. Background technique [0002] Hepatitis B is still one of the most serious public health problems in the world. Hepatitis B ranks seventh among the top 10 causes of death in the world. According to the World Health Organization, about 30% of the world's population, or 2 billion people, have serological evidence of hepatitis B virus infection, of which 350 million people are chronically infected, and at least 1 million people die from HBV infection every year. liver failure, cirrhosis, and hepatocellular carcinoma. And China is the recognized hepatitis B big country in the world, and the carrier of hepatitis B virus of our country reaches 120 million people, and chronic hepatitis B patient has 20 million, and the number of death because of this is nearly 500,000 every year. The prevention, diagnosis and treatment of...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/686C12Q1/70C12Q2545/101C12Q2561/101C12Q2545/114
Inventor 李望丰关尔鑫赵世源蔡一荣王丹肖樊周凯任旭
Owner 东北制药集团辽宁生物医药有限公司
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