口蹄疫O型病毒二温式RT-PCR检测方法的建立及初步应用

doi:10.16431/j.cnki.1671-7236.2017.02.002

Abstract

Abstract:

According to the gene sequences analysis of foot and mouth disease virus (FMDV) in GenBank,a pair of specific primers was designed in the conserved sequence of type O FMDV P1 gene. The reaction parameters were optimized using the uniform design method to develop a two-temperature RT-PCR method for detection of type O FMDV.The results of sensitivity and specificity showed that the two-temperature RT-PCR method was only specific for type O FMDV without amplification of the other viruses. The amplified fragment was same with the expected length.The cloning and sequencing results revealed that the sequence of amplified fragment had 100% simililarity to the target sequence,and the minimum detection quantity was 1.665 pg/μL,the effective detection rate was consistent with the three step RT-PCR sensitivity test results. 54 taper toxicity test pigs were detected,and positive identification results and three-step PCR results was consistent.Compared with the three-step PCR,it could save 20 min.These results indicated that the developed two-temperature RT-PCR for detection of type O FMDV was a kind of accurate,rapid,specific and sensitive detection method.

Key words: type O foot and mouth disease virus; two-temperature RT-PCR; detection method

CLC Number:

S855.3

YAO Huai-bing, ZHAO Yi, LIU Hong, ZHANG Yi, REN Fang, HUANG Jiong. Development and Application of Two-temperature RT-PCR for Detectionof Type O Foot and Mouth Disease Virus[J]. , 2017, 44(2): 311-318.

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Development and Application of Two-temperature RT-PCR for Detectionof Type O Foot and Mouth Disease Virus

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