Primers, probes and kits for detection of hepatitis A and hepatitis E virus

A technology for hepatitis E virus and hepatitis A virus, which is applied in the field of primer probe sets for detecting hepatitis A and hepatitis E virus, and can solve problems such as inability to detect hepatitis A and hepatitis E.

Active Publication Date: 2021-10-22
北京卓诚惠生生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of this disclosure is to solve the defects in the prior art that hepatitis A and hepatitis E cannot be detected rapidly, sensitively and specifically, and provide a detection primer probe and kit for hepatitis A and hepatitis E virus, And establish a rapid, sensitive, specific and simple detection method for hepatitis A and hepatitis E virus based on Recombinase Polymerase Amplification (RPA) technology

Method used

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  • Primers, probes and kits for detection of hepatitis A and hepatitis E virus
  • Primers, probes and kits for detection of hepatitis A and hepatitis E virus
  • Primers, probes and kits for detection of hepatitis A and hepatitis E virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] This example is used to illustrate Hepatitis A and Hepatitis E virus-specific primer probe verification tests.

[0063] (1) Synthesis of primers and probes

[0064] A reagent company was commissioned to synthesize primer-probe sets 1-3 shown in Table 2.

[0065] Table 2

[0066]

Embodiment 2

[0067] Embodiment 2 specificity verification:

[0068] Select the following samples as simulated interference samples: Norovirus (from National CDC, 2001), Hepatitis B virus (from National CDC, 2005), Hepatitis C virus (from National CDC, 2006), Salmonella (from National Culture Collection Center, CVCC3949), Shigella (from National Culture Collection, CVCC3914), Staphylococcus aureus (from National Culture Collection, CVCC1882), Bacillus cereus (from National Culture Collection, CVCC1782), Listeria monocytogenes (from the National Culture Collection, CVCC1345), Yersinia enterocolitica (from the National Culture Collection, CVCC1365), Enterobacter sakazakii (from the National Culture Collection , CVCC1768), Escherichia coli (from National Culture Collection, CVCC2356), Vibrio cholerae (from National Culture Collection, CVCC3467), Escherichia coli O157 (from National Culture Collection, CVCC7389), Aeromonas hydrophila The nucleic acid of bacteria (from the National Culture Coll...

Embodiment 3

[0072] Embodiment 3 minimum detection limit verification:

[0073] Use a concentration of 10 4 The copies / μL of hepatitis A virus and hepatitis E virus nucleic acid were mixed in equal proportions as a template, and the gradient was diluted to 10 3 copies / μL, 10 2 copies / μL, 10 copies / μL, L, and 1 copy / μL. The verification was carried out according to the reaction procedure of the above reaction system. System preparation and RPA reaction conditions are the same as those in Example 2 for specificity evaluation. In the reaction system, the concentration of the virus was 5×10 4 copy / system, 5×10 3 copy / system, 5×10 2 copies / system, 50 copies / system, 10 copies / μL and 5 copies / system.

[0074] Judgment of RPA reaction results: blank control, IAC and negative control are established, otherwise the experimental results will be considered invalid; if the FAM channel has an amplification curve, the primer pair of SEQ ID NO.1-2 and the probe of SEQ ID NO.5 can be detected If th...

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Abstract

The present disclosure discloses a primer probe set, a kit and a detection method for detection of hepatitis A and hepatitis E by recombinant enzyme polymerase amplification, including primers having the nucleotide sequence shown in SEQ ID NO.1-4, and, Containing the probe of the nucleotide sequence shown in SEQ ID NO.5-6. The present disclosure also provides a kit for detection of hepatitis A and hepatitis E viruses by amplifying recombinant enzyme polymerase, said kit including the primer probe set described in the present disclosure. Through the above technical solution, the present disclosure significantly improves the sensitivity, specificity and simplicity of detecting hepatitis A and hepatitis E viruses.

Description

technical field [0001] The disclosure relates to the field of biotechnology, in particular to a primer probe set, a kit and a detection method for detecting hepatitis A and hepatitis E viruses. Background technique [0002] Hepatitis is a serious disease that is widely prevalent in the world and seriously endangers human health and even life. At present, it is mainly divided into hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, and hepatitis G. According to the route of transmission, it can be divided into two categories. Among them, the main routes of transmission of hepatitis B, C, D, and G are blood transmission, iatrogenic transmission, mother-to-child transmission, and sexual transmission. Among them, blood transmission and sexual transmission Transmission is the most common route. However, hepatitis A and E are mainly transmitted through the fecal-oral route, and they are mostly acute infectious diseases on a large scale. [0003] Hepatitis A and Hepa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/706C12Q1/707C12Q2521/107C12Q2521/507C12Q2522/101C12Q2563/107Y02A50/30
Inventor 王雷林笑冬张志强
Owner 北京卓诚惠生生物科技股份有限公司
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