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首页> 外文期刊>Journal of Clinical Microbiology >Isolation and Characterization of Porcine Deltacoronavirus from Pigs with Diarrhea in the United States
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Isolation and Characterization of Porcine Deltacoronavirus from Pigs with Diarrhea in the United States

机译：美国腹泻猪的猪三角洲冠状病毒的分离与鉴定

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Porcine deltacoronavirus (PDCoV) is a novel coronavirus that causes diarrhea in nursing piglets. Following its first detection in the United States in February 2014, additional PDCoV strains have been identified in the United States and Canada. Currently, no treatments or vaccines for PDCoV are available. In this study, U.S. PDCoV strain OH-FD22 from intestinal contents of a diarrheic pig from Ohio was isolated in swine testicular (ST) and LLC porcine kidney (LLC-PK) cell cultures by using various medium additives. We also isolated PDCoV [OH-FD22(DC44) strain] in LLC-PK cells from intestinal contents of PDCoV OH-FD22 strain-inoculated gnotobiotic (Gn) pigs. Cell culture isolation and propagation were optimized, and the isolates were serially propagated in cell culture for >20 passages. The full-length S and N genes were sequenced to study PDCoV genetic changes after passage in Gn pigs and cell culture (passage 11 [P11] and P20). Genetically, the S and N genes of the PDCoV isolates were relatively stable during the first 20 passages in cell culture, with only 5 nucleotide changes, each corresponding to an amino acid change. The S and N genes of our sequenced strains were genetically closely related to each other and to other U.S. PDCoV strains, with the highest sequence similarity to South Korean strain KNU14-04. This is the first report describing cell culture isolation, serial propagation, and biological and genetic characterization of cell-adapted PDCoV strains. The information presented in this study is important for the development of diagnostic reagents, assays, and potential vaccines against emergent PDCoV strains.
机译：猪三角冠状病毒（PDCoV）是一种新型冠状病毒，可引起哺乳仔猪腹泻。自2014年2月在美国首次发现该病毒以来，在美国和加拿大又发现了其他PDCoV毒株。当前，没有针对PDCoV的治疗或疫苗。在这项研究中，通过使用各种培养基添加剂在猪睾丸（ST）和LLC猪肾（LLC-PK）细胞培养物中分离了来自俄亥俄州腹泻猪肠道成分的美国PDCoV株OH-FD22。我们还从接种PDCoV OH-FD22株的生成生猪（Gn）猪的肠内容物中分离了LLC-PK细胞中的PDCoV [OH-FD22（DC44）株]。优化了细胞培养物的分离和繁殖，并将分离物在细胞培养物中连续繁殖> 20代。对全长的S和N基因进行测序，以研究在Gn猪中传代和细胞培养后PDCoV的遗传变化（传代11 [P11]和P20）。从基因上讲，PDCoV分离株的S和N基因在细胞培养的前20代中相对稳定，只有5个核苷酸变化，每个对应于氨基酸变化。我们测序菌株的S和N基因彼此之间以及与其他美国PDCoV菌株在遗传上密切相关，与韩国菌株KNU14-04的序列相似性最高。这是第一份描述细胞培养的PDCoV菌株的细胞培养分离，连续繁殖以及生物学和遗传学特征的报告。这项研究中提供的信息对于开发针对新兴PDCoV株的诊断试剂，分析方法和潜在疫苗非常重要。

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