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Isolation and characterization of a new porcine epidemic diarrhea virus variant that occurred in Korea in 2014

机译:2014年在韩国发生的新的猪流行性腹泻病毒变异株的分离和鉴定

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摘要

Outbreaks of porcine epidemic diarrhea (PED) have resulted in significant economic losses in the swine industry, and another PED outbreak occurred in 2014 in Korea. Isolating and culturing PED virus (PEDV) allow investigations into its pathogenesis and the development of vaccines and diagnostic assays. In this study, we successfully isolated two PEDV isolates (QIAP1401 and QIAP1402) from naturally infected piglets at Jeju-do, Korea. Viral propagation was confirmed in Vero cells based on cytopathic effect, immunofluorescence assay, reverse transcription-polymerase chain reaction, and electron microscopic analyses. The QIAP401 isolate propagated well in Vero cells for 70 passages, with titers of 106.5 to 107.0 50% tissue culture infectious dose/mL, which increased gradually with passaging. The nucleotide and amino acid sequences of the QIAP1401 isolate were determined and compared with those of other PEDV isolates. The QIAP1401 isolate was determined to be closely related to the USA/Minnesota271/2014 strain (> 99.9% nucleotide similarity) that was isolated in the USA in 2014. Phylogenetic analysis based on several PEDV genes suggested that a new PEDV variant is circulating in the Korean swine industry, with 93.08% similarity to the SM98 strain isolated in 1998. In addition, the QIAP1401 strain showed strong virulence in 3-day-old piglets and 11-week-old growing pigs.
机译:猪流行性腹泻(PED)的爆发已导致养猪业的重大经济损失,2014年韩国又发生了一次PED爆发。分离和培养PED病毒(PEDV)可以对其发病机理进行研究,并开发疫苗和进行诊断分析。在这项研究中,我们成功地从韩国济州岛的自然感染仔猪中分离出两个PEDV分离株(QIAP1401和QIAP1402)。根据细胞病变效应,免疫荧光测定,逆转录-聚合酶链反应和电子显微镜分析,证实了Vero细胞中的病毒繁殖。 QIAP401分离株在Vero细胞中繁殖良好,可传70代,滴度为10 6.5 至10 7.0 50%组织培养感染剂量/ mL,并随传代而逐渐增加。确定了QIAP1401分离株的核苷酸和氨基酸序列,并将其与其他PEDV分离株的核苷酸和氨基酸序列进行了比较。确定QIAP1401分离株与2014年在美国分离的USA / Minnesota271 / 2014菌株(> 99.9%核苷酸相似性)密切相关。基于几个PEDV基因的系统发育分析表明,新的PEDV变异体正在循环中。韩国养猪业与1998年分离出的SM98株具有93.08%的相似性。此外,QIAP1401株在3日龄仔猪和11周龄成年猪中显示出强毒力。

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