Belsham G J, Rasmussen T B, Normann P, Vaclavek P, Strandbygaard B, Bøtner A
DTU National Veterinary Institute, Technical University of Denmark, Lindholm, Kalvehave, Denmark.
Department of Virology, State Veterinary Institute Jihlava, Jihlava, Czech Republic.
Transbound Emerg Dis. 2016 Dec;63(6):595-601. doi: 10.1111/tbed.12579. Epub 2016 Sep 12.
During a severe outbreak of diarrhoea and vomiting in a pig herd in Central Eastern Europe, faecal samples were tested positive for porcine epidemic diarrhoea virus (PEDV) and negative for transmissible gastroenteritis virus (TGEV) using a commercial RT-qPCR assay that can detect both of these coronaviruses. However, further analyses, using other TGEV- and PEDV-specific RT-qPCR assays, provided results inconsistent with infection by either of these viruses. Sequencing of an amplicon (ca. 1.6 kb), generated by an RT-PCR specific for the PEDV S-gene, indicated a very close similarity (ca. 99% identity) to recently described chimeric viruses termed swine enteric coronaviruses (SeCoVs). These viruses (with an RNA genome of ca. 28 kb) were first identified in Italy in samples from 2009 but have not been detected there since 2012. A closely related virus was detected in archived samples in Germany from 2012, but has not been detected subsequently. Building on the initial sequence data, further amplicons were generated and over 9 kb of sequence corresponding to the 3'-terminus of the new SeCoV genome was determined. Sequence comparisons showed that the three known SeCoVs are ≥98% identical across this region and contain the S-gene and 3a sequences from PEDV within a backbone of TGEV, but the viruses are clearly distinct from each other. It is demonstrated, for the first time, that pigs from within the SeCoV-infected herd seroconverted against PEDV but tested negative in a TGEV-specific ELISA that detects antibodies against the S protein. These results indicate that SeCoV is continuing to circulate in Europe and suggest it can cause a disease that is very similar to PED. Specific detection of the chimeric SeCoVs either requires development of a new diagnostic RT-qPCR assay or the combined use of assays targeting the PEDV S-gene and another part of the TGEV genome.
在中东欧一个猪群中发生严重腹泻和呕吐疫情期间,使用一种可检测这两种冠状病毒的商业逆转录定量聚合酶链反应(RT-qPCR)检测方法,粪便样本检测出猪流行性腹泻病毒(PEDV)呈阳性,而传染性胃肠炎病毒(TGEV)呈阴性。然而,使用其他TGEV和PEDV特异性RT-qPCR检测方法进行的进一步分析,得到的结果与这两种病毒的感染情况不一致。通过针对PEDV S基因的RT-PCR产生的一个扩增子(约1.6 kb)测序显示,与最近描述的称为猪肠道冠状病毒(SeCoV)的嵌合病毒有非常高的相似性(约99%的同一性)。这些病毒(RNA基因组约28 kb)于2009年在意大利的样本中首次被鉴定出来,但自2012年以来在那里未再检测到。2012年在德国的存档样本中检测到一种密切相关的病毒,但随后未再检测到。基于初始序列数据,生成了更多扩增子,并确定了对应于新SeCoV基因组3'末端的超过9 kb的序列。序列比较表明,三种已知的SeCoV在该区域的同一性≥98%,并且在TGEV的主干内包含来自PEDV的S基因和3a序列,但这些病毒彼此明显不同。首次证明,来自感染SeCoV猪群的猪针对PEDV发生了血清转化,但在检测针对S蛋白抗体的TGEV特异性酶联免疫吸附测定(ELISA)中呈阴性。这些结果表明SeCoV仍在欧洲传播,并提示它可引起与PED非常相似的疾病。嵌合SeCoV的特异性检测要么需要开发一种新的诊断RT-qPCR检测方法,要么需要联合使用针对PEDV S基因和TGEV基因组另一部分的检测方法。