中文摘要：

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1.51×10^4、1.19×10^3、1.26×10^4 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。

英文摘要：

Three pairs of primers were designed separately based on the published porcine kobuvirus （PKV） 3D gene,porcine astrovirus（PAstV） ORF2 gene and porcine torovirus（PToV） M gene sequences. Through the optimization of the reaction system and conditions, a multiplex reverse transcription polyme- rase 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 × 10^4 copies/μL, 1.19 × 10^3 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 consis- tent 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.