PDF(Pubmed)
Abstract：
Porcine respiratory coronavirus (PRCoV), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), and pseudorabies virus (PRV) are significant viruses causing respiratory diseases in pigs. Sick pigs exhibit similar clinical symptoms such as fever, cough, runny nose, and dyspnea, making it very difficult to accurately differentially diagnose these diseases on site. In this study, a quadruplex one-step reverse-transcription real-time quantitative PCR (RT-qPCR) for the detection of PRCoV, PRRSV, SIV, and PRV was established. The assay showed strong specificity, high sensitivity, and good repeatability. It could detect only PRCoV, PRRSV, SIV, and PRV, without cross-reactions with TGEV, PEDV, PRoV, ASFV, FMDV, PCV2, PDCoV, and CSFV. The limits of detection (LODs) for PRCoV, PRRSV, SIV, and PRV were 129.594, 133.205, 139.791, and 136.600 copies/reaction, respectively. The intra-assay and inter-assay coefficients of variation (CVs) ranged from 0.29% to 1.89%. The established quadruplex RT-qPCR was used to test 4909 clinical specimens, which were collected in Guangxi Province, China, from July 2022 to September 2023. PRCoV, PRRSV, SIV, and PRV showed positivity rates of 1.36%, 10.17%, 4.87%, and 0.84%, respectively. In addition, the previously reported RT-qPCR was also used to test these specimens, and the agreement between these methods was higher than 99.43%. The established quadruplex RT-qPCR can accurately detect these four porcine respiratory viruses simultaneously, providing an accurate and reliable detection technique for clinical diagnosis.
摘要：
猪呼吸道冠状病毒（PRCoV），猪繁殖与呼吸综合征病毒(PRRSV),猪流感病毒（SIV），伪狂犬病病毒（PRV）是引起猪呼吸道疾病的重要病毒。病猪表现出类似的临床症状，如发烧，咳嗽,流鼻涕,呼吸困难,这使得很难在现场准确鉴别诊断这些疾病。在这项研究中,四重一步逆转录实时定量PCR(RT-qPCR)检测PRCoV,PRRSV,SIV,PRV成立。该测定法显示出强特异性，高灵敏度,和良好的重复性。它只能检测到PRCoV，PRRSV,SIV,和PRV,没有与TGEV的交叉反应，PEDV,PRoV,ASFV,FMDV,PCV2,PDCoV,CSFVPRCoV的检测限（LOD），PRRSV,SIV,PRV为129.594、133.205、139.791和136.600拷贝/反应，分别。测定内和测定间变异系数(CV)范围为0.29%至1.89%。建立的四重RT-qPCR用于检测4909个临床标本，这些都是在广西收集的，中国,从2022年7月到2023年9月。PRCoV,PRRSV,SIV,PRV的阳性率为1.36%，10.17%，4.87%,和0.84%，分别。此外,先前报道的RT-qPCR也用于测试这些样本，这些方法之间的一致性高于99.43%。建立的四重RT-qPCR可以同时准确检测这四种猪呼吸道病毒，为临床诊断提供准确可靠的检测技术。
