Primers, probes and kits for simultaneous detection of human herpesvirus types 6 and 7

A technology for simultaneous detection of human herpes virus, which is applied in the field of probes and kits, type 7 primers, and simultaneous detection of human herpes virus 6, can solve the problems of high false positive rate, achieve good specificity, reduce false positive rate, The effect of high sensitivity

Active Publication Date: 2020-02-18
武汉光谷创鑫医药孵化服务有限公司
View PDF8 Cites 0 Cited by
  • 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 7 with high sensitivity and high specificity

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primers, probes and kits for simultaneous detection of human herpesvirus types 6 and 7
  • Primers, probes and kits for simultaneous detection of human herpesvirus types 6 and 7
  • Primers, probes and kits for simultaneous detection of human herpesvirus types 6 and 7

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The present embodiment provides a set of primers and fluorescent probes for simultaneous detection of human herpesvirus 6 and 7. The reorganization contains two pairs of primers and two probes. The sequence is 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, the nucleotide sequence of which is 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 quencher group BHQ;

[0037] (2) Forward primer HHV7-F, the nucleotide sequence of which is shown in SEQ ID NO.4;

[0038] Reverse primer HHV7-R, the nucleotide sequence of which is shown in SEQ ID NO.5;

[0039] 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 JOE, and the 3' end quencher group BHQ;

[0040] The above primers...

Embodiment 2

[0042] This embodiment provides a fluorescent quantitative PCR kit for simultaneous detection of human herpesvirus 6 and 7

[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 a commercially available DNA extraction kit, or it can be prepared by itself.

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

[0049] 3) The reaction system of double fluorescent 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-7 primer 0.29μmol / L ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention discloses primers, probes and kit for synchronously detecting human herpesvirus-6 and human herpesvirus-7. According to the primers, the probes and the kit, through selecting a novel target gene and redesigning the primers and the probes, the reaction system is optimized, the false positive incidence of clinical synchronous detection on human herpesviruses is substantially lowered, the sensitivity is high, the specificity is good, and the time for detection is shorter, so that the clinical application and popularization of the technology 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 synchronous detection of human herpes virus types 6 and 7. Background technique [0002] Existing studies have confirmed that human herpesvirus (human herpesvirus, HHV)-6, -7, -8 can be transmitted through blood transfusion, and invade human peripheral blood mononucleated cells CD4 + T cells, which subsequently enter a latent state and cause persistent infection, are activated to cause disease in immunocompromised and neurologically diseased individuals. [0003] For herpes virus types 6, 7, and 8, a multiplex fluorescent quantitative PCR method is provided in the prior art, and U67, U36, and ORF65 are selected as target genes of HHV-6, HHV-7, and HHV-8 respectively. The above three viruses can be detected at one time, and the sensitivity is high, but its defect is that the false positive rate is high in use, especially for HHV-6 an...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
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 武汉光谷创鑫医药孵化服务有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap