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Primer, probe and kit for simultaneously detecting human herpesvirus 6 and 8

A technology for simultaneous detection of human herpesviruses, applied to primers, probes and kits of type 8, and simultaneous detection of human herpesvirus 6 fields, can solve the problem of high false positive rate, and achieve good specificity, high sensitivity, and optimized reaction. effect of the system

Inactive Publication Date: 2016-12-21
武汉百格资产管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the defects of high false positive rate of existing herpes virus detection methods, and provide a kind of primers, probes and kits for simultaneous detection of human herpes virus 6 and 8 with high sensitivity and high specificity

Method used

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  • Primer, probe and kit for simultaneously detecting human herpesvirus 6 and 8
  • Primer, probe and kit for simultaneously detecting human herpesvirus 6 and 8
  • Primer, probe and kit for simultaneously detecting human herpesvirus 6 and 8

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The present embodiment provides a set of primers and fluorescent probes for synchronous detection of human herpesvirus types 6 and 8, the reorganization contains three pairs of primers and three probes, and the sequences are as follows:

[0034] (1) Forward primer HHV6-F, the nucleotide sequence of which is shown in SEQ ID NO.1;

[0035] Reverse primer HHV6-R, its nucleotide sequence is as shown in SEQ ID NO.2;

[0036] Fluorescent probe HHV6-P, the nucleotide sequence of which is shown in SEQ ID NO.3, the 5' end of the fluorescent probe is labeled with FAM, and the 3' end is quenched by BHQ;

[0037] (2) forward primer HHV8-F, its nucleotide sequence is as shown in SEQ ID NO.4;

[0038] Reverse primer HHV8-R, its nucleotide sequence is as shown in SEQ ID NO.5;

[0039] The fluorescent probe HHV8-P, the nucleotide sequence of which is shown in SEQ ID NO. 6, is labeled with Cy5 at the 5' end of the fluorescent probe and a quenching group BHQ at the 3' end.

[0040] Th...

Embodiment 2

[0042] This embodiment provides a fluorescence quantitative PCR kit for simultaneous detection of human herpesvirus 6 and 8

[0043] 1. Kit composition and preparation

[0044] 1) Primer and fluorescent probe set

[0045] Same as Example 1

[0046] 2) DNA extraction solution

[0047] The DNA extraction solution can use commercially available DNA extraction kits, or can be prepared by yourself.

[0048] In this example, the formula of DNA extract: prepare 200mmol / L NaCl, 100mmol / L Tris-HCl, 1% SDS, 50mmol / L EDTA, adjust pH to 8.5, 3mg / ml proteinase K (add before use)

[0049] 3) The reaction system of double fluorescence quantitative PCR is as follows:

[0050] Reagent

Final concentration or content per unit volume

PCR 10×buffer

5μl

MgCl 2

4mmol / L

dNTP (including dUTP)

0.48mmol / L

Tag DNA polymerase

0.5U

UNGase

0.2U

HHV-6 primer

0.29μmol / L

HHV-6 probe

0.24μmol / L

HHV-8 primer

...

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PUM

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Abstract

The invention discloses a primer, probe and kit for simultaneously detecting human herpesvirus 6 and 8. The primer, probe and kit has the advantages that the primer and the probe are redesigned by selecting new target genes, a reaction system is optimized, the false positive rate of simultaneous human herpesvirus detection is lowered greatly, the primer, probe and kit is high in sensitivity, good in specificity and short in detection time, and clinical application and popularization can be achieved.

Description

technical field [0001] The invention relates to the technical field of herpes virus detection, in particular to a primer, a probe and a kit for simultaneous detection of human herpes virus types 6 and 8. Background technique [0002] Existing studies have confirmed that human herpesvirus (human herpesvirus, HHV)-6, -7, -8 can be transmitted through blood transfusion, and invade the CD4 of human peripheral blood mononucleated cells after primary infection. + T cells, which subsequently enter a latent state, cause persistent infection, and become pathogenic when activated in immunocompromised and neurologically ill individuals. [0003] For herpes virus types 6, 7 and 8, a multiplex fluorescence quantitative PCR method is provided in the prior art, and U67, U36 and ORF65 are selected as the target genes of HHV-6, HHV-7 and HHV-8 respectively. The above three viruses can be detected at one time with high sensitivity, but the defect is that the false positive rate in use is hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/705C12Q1/6851C12Q2600/16C12Q2531/113C12Q2537/143C12Q2545/113C12Q2563/107
Inventor 孙莉军李卫赵友云郑毅王汉敏蔡望喜
Owner 武汉百格资产管理有限公司
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