Primer-probe assembly for detecting cytomegalovirus, EB (epstein-barr) virus and adenovirus

A cytomegalovirus, Epstein-Barr virus technology, applied in microorganism-based methods, microorganisms, recombinant DNA technology, etc., can solve the problems of unfavorable acute diagnosis, cumbersome operation, long time-consuming, etc., to shorten working time, overcome easy pollution, overcome limited effect

Inactive Publication Date: 2016-12-07
ZHEJIANG UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

The cell culture and serological detection of the virus are cumbersome and time-consuming, which is not conducive to acute diagnosis. When the viral load is low, it is easy to cause missed detection

Method used

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  • Primer-probe assembly for detecting cytomegalovirus, EB (epstein-barr) virus and adenovirus
  • Primer-probe assembly for detecting cytomegalovirus, EB (epstein-barr) virus and adenovirus
  • Primer-probe assembly for detecting cytomegalovirus, EB (epstein-barr) virus and adenovirus

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Embodiment Construction

[0040] 1. Design of primers and probes: through comparative analysis of all known cytomegalovirus, Epstein-Barr virus, and adenovirus gene sequences, select highly conserved segments and design multiple pairs of primers and probes. The length of the primers is average It is about 20 bases. The optimal primer-probe sequence combination is as follows:

[0041] (1) CMV primers and probes:

[0042] Upstream primer CMV F: GCAAATATGGAAACATACGTGAACA

[0043] Downstream primer CMV R: GCACCCATATTGTWAGTGATGCA

[0044] CMV Probe: CTTCACGAAGGTCTCACATACACAGCWG-FAM-BHQ

[0045] (2) Epstein-Barr virus primers and probes:

[0046]Upstream primer EB F: GCCACGGTGGGGTTTCTAAACTT

[0047] Downstream primer EB R:GCCCCAGTGGTCTTACATGCACATC

[0048] Probe EB Probe: TGCACCAGACCCGGGCTCAGGTACTCCGA-HEX-BHQ1

[0049] (3) Adenovirus primers and probes:

[0050] Upstream primer ADV F: GTTGTCAATGTCTTAATTCGTATCAATAATT

[0051] Downstream primer ADV R:GTAGCCTMCCTTCGGCACCTAA

[0052] Probe ADV Probe: T...

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Abstract

The invention relates to primers and probes for detection of various respiratory viruses and provides a primer-probe assembly for detecting cytomegalovirus, EB (epstein-barr) virus and adenovirus. Sequences of a probe and a primer pair for detecting the cytomegalovirus, the EB virus and the adenovirus are as shown in SEQ ID NO:1-9. The invention further provides a kit for simultaneous detection of the cytomegalovirus, the EB virus and the adenovirus. By combination of multiplex PCR (polymerase chain reaction) and Taqman fluorescent probe technique, simultaneous detection of various pathogens possibly causing acute respiratory infection can be realized in one-time detection, and quickness in diagnosis and treatment guide is realized. In addition, shortening of operation time and reduction of raw material consumption are realized, and high sensitivity and specificity, effectiveness in solving of PCR pollution, high automation degree and the like are achieved.

Description

technical field [0001] The invention relates to primers and probes for detecting various respiratory viruses. In particular, the present invention is a primer and probe capable of specific detection of cytomegalovirus, Epstein-Barr virus and adenovirus at the same time. Background technique [0002] More than 90% of acute respiratory infections are caused by viruses, among which respiratory viruses are the most common. Common respiratory viruses include: influenza A virus, influenza B virus, parainfluenza virus, cytomegalovirus, respiratory syncytial virus, adenovirus, Epstein-Barr virus, etc. Cytomegalovirus (Cytomegaoviyns, CMV) has a typical herpes virus morphology and is highly species-specific to hosts or cultured cells. Human cytomegalovirus can only infect humans and proliferate only in human fibroblasts. Children with CMV infection mostly manifest as asymptomatic infection, only a few are symptomatic infection, and most of them occur in children with congenital and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/70C12Q1/686C12Q1/701C12Q2600/16C12Q2537/143C12Q2561/101
Inventor 张晨美
Owner ZHEJIANG UNIV
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