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Method and kit for detecting poliomyelitis viruses in food in real time through fluorescent RT-PCR

A polio, kit technology, applied in the fields of molecular biology and nucleic acid detection

Active Publication Date: 2012-12-26
PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, my country only conducts routine bacterial detection of imported and exported food, and there is no corresponding method to detect poliovirus in imported and exported food
However, there are currently no assays for the rapid and easy detection and identification of poliovirus
[0008] In summary, in view of the fact that there is no real-time fluorescent RT-PCR method for the detection of poliovirus in food with good specificity and high sensitivity

Method used

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  • Method and kit for detecting poliomyelitis viruses in food in real time through fluorescent RT-PCR
  • Method and kit for detecting poliomyelitis viruses in food in real time through fluorescent RT-PCR
  • Method and kit for detecting poliomyelitis viruses in food in real time through fluorescent RT-PCR

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Experimental program
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Effect test

Embodiment 1

[0100] 1 Materials and methods

[0101] 1.1 Materials and reagents

[0102] Serum inactivated vaccine suspensions of poliovirus types I, II, and III were obtained from the University of California, USA, and inactivated coxsackievirus types A9, A16, B2, B3, and B5 and stool samples containing norovirus types GII were purchased from China CDC Institute for Viral Prevention and Control.

[0103] PBS buffer: 0.14mol / L NaCl, 0.003mol / L KCl, 0.00175mol / L KH 2 PO 4 , 0.01mol / L Na 2 HPO 4 , pH 7.5, Shanghai Sangon Bioengineering Co., Ltd.; glycine buffer: 0.1mol / L glycine, 0.3mol / L NaCl, pH 9.5; PEG 8000 solution: 16% PEG 8000, 0.525mol / L NaCl; TIANamp virus 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.

[0104] 1.2 Primers and probes

[0105] Compa...

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Abstract

The invention relates to a method for detecting poliomyelitis viruses in food in real time through fluorescent reverse transcription-polymerase chain reactions (RT-PCR). Particularly, the invention discloses a method for detecting the poliomyelitis viruses. In the method, a polymerase chain reaction is performed in a polymerase reaction system which contains a specific primer pair for amplifying the poliomyelitis viruses and a specific probe of the poliomyelitis viruses. The invention also provides a corresponding kit. By using the method and the kit, the poliomyelitis viruses can be detected and identified sensitively, simply and conveniently.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection. Specifically, the invention relates to a real-time fluorescent PCR detection method and kit for poliovirus in foods such as shellfish, vegetables and fruits, and water. Background technique [0002] In recent years, outbreaks of diseases caused by eating virus-contaminated food have been frequently reported around the world, among which diseases caused by enteroviruses accounted for more than 67%. [0003] As a typical pathogenic enterovirus, although the wild strain of poliovirus has been basically eliminated all over the world, due to the widespread use of poliovirus vaccines, its mutant strains have caused many virus outbreaks in recent years event. According to the World Health Organization, the number of cases of vaccine-derived poliovirus infection rose to 80 in 2008, compared with nearly 1,800 cases of wild-type virus infection. In order to strength...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 李想潘良文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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