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Real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method and kit for Coxsackie virus

A coxsackie virus and kit technology, applied in the field of molecular biology and nucleic acid detection, can solve the problem of limited group resolution, lack of serotype coxsackie virus high sensitivity, high specificity, flexible and simple detection method, poor sensitivity And other issues

Inactive Publication Date: 2012-01-11
PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Description
  • Claims
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Problems solved by technology

Previous studies were mostly based on Taqman-FAM-TAMRA probes, for example: Tana (2008) established a multiplex real-time fluorescent PCR method for Coxsackievirus A16 and Enterovirus 71 [5] However, the method used to detect A16 virus is a fluorescent hybridization probe, which has higher requirements for the real-time fluorescent PCR system, and not all real-time fluorescent PCR instruments can implement the reaction
Fluorescent hybridization probes require 2 adjacent probes, which increases cost, and one of the probes is labeled with the LC Red 750 fluorophore, which has limited resolution and poor sensitivity
Therefore, this method is not conducive to popularization and application
[0007] In this field, there is still a lack of highly sensitive, highly specific, flexible, simple, and highly applicable detection methods for a single serotype of Coxsackieviruses, and there is even a lack of detection techniques for rapid detection and identification of various serotypes of Coxsackieviruses
[0008] In summary, in view of the fact that there is no real-time fluorescent RT-PCR detection method with good specificity and high sensitivity for Coxsackie virus

Method used

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  • Real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method and kit for Coxsackie virus
  • Real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method and kit for Coxsackie virus
  • Real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method and kit for Coxsackie virus

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Experimental program
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Effect test

Embodiment 1

[0117] 1 Materials and methods

[0118] 1.1 Materials and reagents

[0119] Inactivated coxsackievirus types A9, A16, B2, B3, and B5 and norovirus-containing GII stool samples were purchased from the Institute of Virology Prevention and Control, Chinese Center for Disease Control and Prevention, and serum types I, II, and III of poliovirus were inactivated. Live vaccine strain suspensions were obtained from the University of California, USA.

[0120] TIANamp Viral RNA Extraction Kit: Tiangen Biotechnology Co., Ltd., Cat.No.SD101; Prime Script TM Reverse transcription kit: Bao Biological Engineering Co., Ltd., Cat.No.DRR037A; Premix Ex Taq TM Real-time fluorescence amplification kit: Bao Biological Engineering Co., Ltd., Cat.No.DRR039A.

[0121] 1.2 Primers and probes

[0122] Search for the cDNA sequences of Coxsackieviruses A9, A16, B2, B3 and B5 in the GenBank database, compare and analyze them, and find out that each serotype is relatively conserved, but it is different...

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Abstract

The invention relates to a real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method and a kit for a Coxsackie virus. Specifically, the invention discloses a method for detecting the Coxsackie virus, which is characterized in that a polymerase chain reaction is carried out in a polymerase reaction system. The polymerase reaction system comprises an amplification Coxsackie virus-contained specific primer pair and a Coxsackie virus specific probe. The invention also provides the corresponding kit. The invention is capable of sensitively, simply and conveniently detecting and verifying the Coxsackie virus.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection. Specifically, the present invention relates to a real-time fluorescent RT-PCR detection method and kit for Coxsackie virus detection. Background technique [0002] Coxsackievirus (Coxsackievirus) belongs to Picornaviridae (Picornaviridae) enterovirus genus, according to its biological characteristics can be divided into two categories, namely A group and B group. Coxsackievirus infection in humans causes herpetic angina (group A) and nonparalytic polio (group B), respectively. According to the survey, 79% of acute fevers accompanied by oropharyngeal herpes and rash are caused by Coxsackie virus A group, and the more common Coxsackie virus A16 type is one of the main pathogens causing hand, foot and mouth disease. Coxsackievirus group B is the main pathogenic microorganism causing viral myocarditis in humans [1] . [0003] In recent years, there have been ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 李想潘良文李俊毅黄一卢钟山张舒亚吕蓉
Owner PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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