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当前位置: 首页 > 文章 > 猪嵴病毒和猪星状病毒及猪环曲病毒多重RT-PCR检测方法的建立及临床应用 中国兽医科学 2015 (7) 661-667
Position: Home > Articles > Establishment and clinical application of a multiplex RT-PCR for detection of porcine kobuvirus,porcine astrovirus and porcine torovirus Chinese Veterinary Science 2015 (7) 661-667

猪嵴病毒和猪星状病毒及猪环曲病毒多重RT-PCR检测方法的建立及临床应用

作  者：
顾凡;周远成;黄剑波;樊毅;赵洲;乔小改;朱玲;刘书亮;徐志文
单  位：
自贡市大安区农牧林业局;四川农业大学食品学院;四川农业大学动物疫病与人类健康四川省重点实验室;四川农业大学动物生物技术中心
关键词：
猪嵴病毒;猪星状病毒;猪环曲病毒;多重RT-PCR
摘  要：
根据猪嵴病毒(PKV)3D基因、猪星状病毒(PAstV)ORF2基因和猪环曲病毒(PToV)M基因的序列,设计了3对引物,通过反应体系和条件的优化,首次建立了检测猪嵴病毒、猪星状病毒和猪环曲病毒的多重RT-PCR。本研究建立的多重RT-PCR对PKV、PAstV和PToV的最低检出限分别为1.51×104、1.19×103、1.26×104 copies/μL,同时也具有较强的特异性和良好的重复性。用该方法检测了40份来自四川省部分猪场的仔猪腹泻病样。结果显示,PKV、PAstV、PToV的阳性检出率分别为75.0%、30.0%和22.5%。与单一RT-PCR检测结果一致。本方法的建立对PKV、PAstV、PToV的鉴别诊断和混合感染检测具有重要的应用价值。
译  名：
Establishment and clinical application of a multiplex RT-PCR for detection of porcine kobuvirus,porcine astrovirus and porcine torovirus
作  者：
GU Fan;ZHOU Yuan-cheng;HUANG Jian-bo;FAN Yi;ZHAO Zhou;QIAO Xiao-gai;ZHU Ling;LIU Shu-liang;XU Zhi-wen;Animal Biotechnology Center,Sichuan Agricultural University;Da’an District Bureau of Agriculture,Animal Husbandry and Forestry;College of Food Science,Sichuan Agricultural University;Key Laboratory of Animal Disease and Human Health of Sichuan Province,Sichuan Agricultural University;
关键词：
porcine kobuvirus;;porcine astrovirus;;porcine torovirus;;multiplex RT-PCR
摘  要：
Three pairs of primers were designed separately based on the published porcine kobuvirus(PKV)3Dgene,porcine astrovirus(PAstV)ORF2gene and porcine torovirus(PToV)M gene sequences.Through the optimization of the reaction system and conditions,a multiplex reverse transcription polymerase chain reaction(multiplex RT-PCR)for detection of PKV,PAstV and PToV was developed for the first time.The lowest detection limits of PKV,PAstV and PToV were proved to be 1.51×104 copies/μL,1.19×103 copies/μL and 1.26×104 copies/μL respectively.And this multiplex RT-PCR method has high specificity and good repeatability.To evaluate the potential application of the multiplex RT-PCR assay for clinical diagnosis,40 samples from piglets with diarrhea in Sichuan Province were obtained and examined.The positive rate of PKV,PAstV and PToV were 75.0%,30.0% and 22.5%,respectively.This result is consistent with the single RT-PCR.The results suggested that this method has the application value in diagnosis or differential diagnosis in PKV,PAstV and PToV mixed infection.

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