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Primers for detecting hepatitis B virus nucleic acids, probe, kit and detection method

A hepatitis B virus and detection method technology, applied in the field of molecular biology, can solve the problems of expensive high-sensitivity diagnostic kits, increased medical and health costs, and high quantitative detection limit, so as to reduce the detection limit, reduce detection costs, increase The effect of sensitivity

Inactive Publication Date: 2019-03-12
SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are a variety of virus detection kits based on fluorescent quantitative PCR technology at home and abroad, but the domestic detection kits have a high quantitative detection limit and poor sensitivity.
However, the high-sensitivity diagnostic kits imported from abroad are too expensive, which increases the cost of medical care and health care.

Method used

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  • Primers for detecting hepatitis B virus nucleic acids, probe, kit and detection method
  • Primers for detecting hepatitis B virus nucleic acids, probe, kit and detection method
  • Primers for detecting hepatitis B virus nucleic acids, probe, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] 1. The samples used in this embodiment include 2 cases of HBV nucleic acid positive nucleic acid solution and 1 case of negative nucleic acid solution.

[0101] 2. Preparation of PCR reaction system

[0102] (1) Take out the PCR reaction solution from the refrigerator in advance to thaw, vortex and mix well after melting completely, and centrifuge instantaneously;

[0103] (2) Take 19 μL of PCR reaction solution and place it in a PCR tube, add 1 μL of nucleic acid sample into it, and mix well with a pipette tip;

[0104] (3) Draw 20 μL of the mixed liquid into the injection hole, draw 20 μL of the droplet generation solution into the reagent well;

[0105] (4) Fix the sample processing device on the centrifuge adapter through the positioning hole, and pay attention to maintaining central symmetry when installing the processing device;

[0106](5) Centrifuge at 3000rpm for 3 minutes, so that the nucleic acid sample and the PCR reaction solution flow into the flow chann...

Embodiment 2

[0118] 1. The samples used in this embodiment include four concentrations of plasmid quantitative reference products containing HBV specific sequences, wherein the plasmid concentrations are A (10copies / μL), B (5copies / μL), C (2copies / μL) and D (1copies / μL).

[0119] 2. Preparation of PCR reaction system

[0120] (1) Take out the PCR reaction solution from the refrigerator in advance to thaw, vortex and mix well after melting completely, and centrifuge instantaneously;

[0121] (2) Take 10 μL of PCR reaction solution and place it in a PCR tube, add 10 μL of nucleic acid sample into it, and mix well with a pipette tip;

[0122] (3) Draw 20 μL of the mixed liquid into the injection hole, draw 20 μL of the droplet generation solution into the reagent well;

[0123] (4) Fix the sample processing device on the centrifuge adapter through the positioning hole, and pay attention to maintaining central symmetry when installing the processing device;

[0124] (5) Centrifuge at 3000rp...

Embodiment 3

[0137] 1. The samples used in this embodiment include 1 case each of human hepatitis C virus (HCV) nucleic acid positive, human herpes virus (EBV) nucleic acid positive, cytomegalovirus (CMV) nucleic acid positive and HBV nucleic acid positive samples.

[0138] 2. Preparation of PCR reaction system

[0139] (1) Take out the PCR reaction solution from the refrigerator in advance to thaw, vortex and mix well after melting completely, and centrifuge instantaneously;

[0140] (2) Take 10 μL of PCR reaction solution and place it in a PCR tube, add 10 μL of nucleic acid sample into it, and mix well with a pipette tip;

[0141] (3) Draw 20 μL of the mixed liquid into the injection hole, draw 20 μL of the droplet generation solution into the reagent well;

[0142] (4) Fix the sample processing device on the centrifuge adapter through the positioning hole, and pay attention to maintaining central symmetry when installing the processing device;

[0143] (5) Centrifuge at 3000rpm for 3...

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Abstract

The invention discloses primers for detecting hepatitis B virus nucleic acids, a probe, a kit and a detection method. A primer pair for detecting hepatitis B virus nucleic acids comprises a first primer and a second primer, and the sequences of the first primer and the second primer are respectively as shown in SEQ ID NO.1 and SEQ ID NO.2. The sequence of the probe is as shown in SEQ ID NO.3. Thekit comprises the primer pair and the probe. The detection method of hepatitis B virus nucleic acids comprises the following steps: mixing a nucleic acid sample to be detected with PCR (Polymerase Chain Reaction) liquid by adopting a sample treatment device, and coating the generated mixed liquid with droplet generation liquid to form droplets which contain single nucleic acids or do not contain nucleic acids; and performing PCR amplification on the droplets by utilizing the kit, and detecting PCR amplification products. The kit disclosed by the invention is low in lower limit of detection, high in sensitivity and good in specificity. The detection method disclosed by the invention is simple and quick, very high in detection efficiency and low in cost.

Description

technical field [0001] The present invention relates to the field of molecular biology technology, more specifically, to the field of nucleic acid technology, in particular to a primer, probe and corresponding kit for highly sensitive detection of hepatitis B virus (HBV) nucleic acid, and the use of the reagent A method for detecting hepatitis B virus nucleic acid with a box. Background technique [0002] Hepatitis B Virus (HBV) belongs to the hepadnaviridae family. It is one of the smallest DNA viruses known to infect humans, and it is also one of the most difficult viruses to cure. HBV can invade the human body through blood, mother-infant, sex, and medical sources, causing liver inflammatory lesions and multiple organ damage. It is an epidemic virus that seriously threatens human health. HBV is a Class B infectious disease in my country, and there were as many as 1 million new cases in 2017 alone. Patients infected with HBV virus may develop chronic hepatitis if they ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/707C12Q2563/159C12Q2563/107
Inventor 张惠丹戴敬赵洪玉节淳皓
Owner SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD
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