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2014年在韩国出现的一种新型猪流行性腹泻病毒变种的分离与鉴定

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

作者信息

Yang Dong-Kun, Kim Ha-Hyun, Lee Seung-Heon, Yoon Soon-Seek, Park Jung-Won, Cho In-Soo

机构信息

Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs, Gimcheon 39660, Korea.

出版信息

J Vet Sci. 2018 Jan 31;19(1):71-78. doi: 10.4142/jvs.2018.19.1.71.

Abstract

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 10 to 10 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.

摘要

猪流行性腹泻(PED)的爆发给养猪业造成了重大经济损失,2014年韩国再次爆发了PED疫情。分离和培养PED病毒(PEDV)有助于对其发病机制进行研究,并开发疫苗和诊断检测方法。在本研究中,我们成功地从韩国济州岛自然感染的仔猪中分离出两株PEDV毒株(QIAP1401和QIAP1402)。基于细胞病变效应、免疫荧光检测、逆转录-聚合酶链反应和电子显微镜分析,在Vero细胞中证实了病毒的增殖。QIAP401毒株在Vero细胞中传代70次,增殖良好,滴度为10至10 50%组织培养感染剂量/毫升,随着传代逐渐增加。测定了QIAP1401毒株的核苷酸和氨基酸序列,并与其他PEDV毒株进行了比较。确定QIAP1401毒株与2014年在美国分离的USA/Minnesota271/2014毒株密切相关(核苷酸相似性>99.9%)。基于多个PEDV基因的系统发育分析表明,一种新的PEDV变体正在韩国养猪业中传播,与1998年分离的SM98毒株相似性为93.08%。此外,QIAP1401毒株对3日龄仔猪和11周龄生长猪表现出很强的毒力。

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