State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
School of Life Sciences, Lanzhou University, Lanzhou, China.
Transbound Emerg Dis. 2020 Mar;67(2):678-685. doi: 10.1111/tbed.13385. Epub 2019 Oct 28.
The major enteric RNA viruses in pigs include porcine epidemic diarrhoea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus A (PRV-A), porcine kobuvirus (PKV), porcine sapovirus (PSaV) and porcine deltacoronavirus (PDCoV). For differential diagnosis, a multiplex RT-PCR method was established on the basis of the N genes of TGEV, PEDV and PDCoV, the VP7 gene of PRV-A, and the polyprotein genes of PKV and PSaV. This multiplex RT-PCR could specifically detect TGEV, PEDV, PDCoV, PRV-A, PKV and PSaV without cross-reaction to any other major viruses circulating in Chinese pig farms. The limit of detection of this method was as low as 10 -10 ng cDNA of each virus. A total of 398 swine faecal samples collected from nine provinces of China between October 2015 and April 2017 were analysed by this established multiplex RT-PCR. The results demonstrated that PDCoV (144/398), PSaV (114/398), PEDV (78/398) and PRV-A (70/398) were the main pathogens, but TGEV was not found in the pig herds in China. In addition, dual infections, for example, PDCoV + PSaV, PDCoV + PRV-A, PRA-V + PSaV and PEDV + PDCoV, and triple infections, for example, PDCoV + PRV-A + PSaV and PEDV + PDCoV + PKV, were found among the collected samples. The multiplex RT-PCR provided a valuable tool for the differential diagnosis of swine enteric viruses circulating in Chinese pig farms and will facilitate the prevention and control of swine diarrhoea in China.
猪的主要肠道 RNA 病毒包括猪流行性腹泻病毒（PEDV）、传染性胃肠炎病毒（TGEV）、猪轮状病毒 A（PRV-A）、猪博卡病毒（PKV）、猪甲病毒（PSaV）和猪德尔塔冠状病毒（PDCoV）。为了进行鉴别诊断，基于 TGEV、PEDV 和 PDCoV 的 N 基因、PRV-A 的 VP7 基因以及 PKV 和 PSaV 的多蛋白基因，建立了一种多重 RT-PCR 方法。该多重 RT-PCR 可特异性检测 TGEV、PEDV、PDCoV、PRV-A、PKV 和 PSaV，与中国猪场流行的其他主要病毒无交叉反应。该方法的检测限低至每种病毒 10 -10 ng cDNA。使用该建立的多重 RT-PCR 分析了 2015 年 10 月至 2017 年 4 月期间从中国 9 个省采集的 398 份猪粪便样本。结果表明，PDCoV（144/398）、PSaV（114/398）、PEDV（78/398）和 PRV-A（70/398）是主要病原体，但在中国猪群中未发现 TGEV。此外，在收集的样本中发现了双重感染，例如 PDCoV + PSaV、PDCoV + PRV-A、PRV-A + PSaV 和 PEDV + PDCoV，以及三重感染，例如 PDCoV + PRV-A + PSaV 和 PEDV + PDCoV + PKV。该多重 RT-PCR 为中国猪场流行的猪肠道病毒的鉴别诊断提供了一种有价值的工具，将有助于中国猪腹泻的预防和控制。
