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Primer, probe and kit for synchronously detecting human herpes virus types 6, 7 and 8

A technology for simultaneous detection of human herpes virus, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of high false positive rate, achieve good specificity, short detection time, and small interaction effects

Active Publication Date: 2016-11-16
湖南光谷创新医疗器械公共服务平台有限公司
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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, 7 and 8 with high sensitivity and high specificity

Method used

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  • Primer, probe and kit for synchronously detecting human herpes virus types 6, 7 and 8
  • Primer, probe and kit for synchronously detecting human herpes virus types 6, 7 and 8
  • Primer, probe and kit for synchronously detecting human herpes virus types 6, 7 and 8

Examples

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Embodiment 1

[0041] The present embodiment provides a set of primers and fluorescent probes for synchronous detection of human herpesvirus types 6, 7, and 8, the reorganization contains three pairs of primers and three probes, and the sequences are as follows:

[0042] (1) Forward primer HHV6-F, the nucleotide sequence of which is shown in SEQ ID NO.1;

[0043] Reverse primer HHV6-R, its nucleotide sequence is as shown in SEQ ID NO.2;

[0044] 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;

[0045] (2) forward primer HHV7-F, its nucleotide sequence is as shown in SEQ ID NO.4;

[0046] Reverse primer HHV7-R, its nucleotide sequence is as shown in SEQ ID NO.5;

[0047] Fluorescent probe HHV7-P, the nucleotide sequence of which is shown in SEQ ID NO.6, the 5' end of the fluorescent probe is labeled with JOE, and the 3' end is a quencher group BHQ;

[0048] (3) fo...

Embodiment 2

[0053] This embodiment provides a fluorescent quantitative PCR kit for simultaneous detection of human herpesvirus types 6, 7, and 8

[0054] 1. Kit composition and preparation

[0055] 1) Primer and fluorescent probe set

[0056] Same as Example 1

[0057] 2) DNA extraction solution

[0058] The DNA extraction solution can use commercially available DNA extraction kits, or can be prepared by yourself.

[0059] In this example, the formula of DNA extraction solution: prepare 200 mmol / L NaCl, 100 mmol / L Tris-HCl, 1% SDS, 50 mmol / L EDTA, adjust pH to 8.5, 3 mg / ml proteinase K (add before use).

[0060] 3) The reaction system of multiplex fluorescence quantitative PCR is as follows:

[0061]

[0062]

[0063] 4) Negative quality control standard: sterile double distilled water containing HSV1 and HSV2.

[0064] 5) Positive quantitative standard:

[0065] HHV-6 (A type U1102 strain, B type Z29 strain), the concentration is 1×10 9 Copies / L;

[0066] HHV-7 (RK strain), t...

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Abstract

The invention discloses a primer, a probe and a kit for synchronously detecting human herpes virus types 6, 7 and 8. By selecting a new target gene, redesigning a primer and a kit, and optimizing a reaction system, the false positive rate of clinical synchronous detection on human herpes viruses is greatly reduced, the primer, the probe and the kit are good in sensitivity, good in specificity and short in detection time, and the technique can be applied to clinical application and popularization.

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 synchronously detecting human herpes virus types 6, 7 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 hi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/705C12Q2600/16C12Q2531/113C12Q2537/143C12Q2545/113
Inventor 赵友云李卫孙莉军郑毅王汉敏蔡望喜
Owner 湖南光谷创新医疗器械公共服务平台有限公司
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