猪德尔塔冠状病毒TJ1株的分离鉴定及生物学特性分析

doi:10.16431/j.cnki.1671-7236.2018.01.028

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (1): 219-224.doi: 10.16431/j.cnki.1671-7236.2018.01.028

猪德尔塔冠状病毒TJ₁株的分离鉴定及生物学特性分析

郑丽^1,2, 李秀丽^1,2, 鄢明华^1,2, 任卫科^1,2, 张蕾^1,2, 路超^1,2, 田向学^1,2, 韩伟³

1. 天津市畜牧兽医研究所, 天津 300381;
2. 天津市畜禽健康养殖技术工程中心, 天津 300381;
3. 天津市西青区乡镇畜牧兽医总站, 天津 300380

收稿日期:2017-07-19 出版日期:2018-01-20 发布日期:2018-01-20
通讯作者: 鄢明华 E-mail:yanmh81971@126.com
作者简介:郑丽(1989-),女,内蒙古呼伦贝尔人,硕士,研究方向:动物分子病原学,E-mail:407126867@qq.com
基金资助:
天津市农业科技转化与推广项目（201301030）；国家重点研发计划资助项目（2016YFD0801002-02）；天津市生猪产业技术体系猪病防控岗位项目（ITTPRS2017003）

Isolation, Identification and Biological Characteristics Analysis of Porcine Deltacoronavirus TJ₁

ZHENG Li^1,2, LI Xiuli^1,2, YAN Minghua^1,2, REN Weike^1,2, ZHANG Lei^1,2, LU Chao^1,2, TIAN Xiangxue^1,2, HAN Wei³

1. Tianjin Institute of Animal Husbandry and Veterinary Science, Tianjin 300381, China;
2. Tianjin Engineering Research Center for Livestock and Poultry Health Breeding, Tianjin 300381, China;
3. Tianjin Xiqing District Township Animal Husbandry and Veterinary Medicine Station, Tianjin 300380, China

Received:2017-07-19 Online:2018-01-20 Published:2018-01-20

摘要/Abstract

摘要：

本研究旨在获得猪德尔塔冠状病毒（porcine deltacoronavirus，PDCoV）分离株，并对其生物学特性进行探讨。将RT-PCR检测PDCoV阳性样品无菌处理后接种猪肾（PK-15）细胞并连续传代培养，通过细胞病变、M基因序列分析及耐热、耐酸等试验对细胞培养物进行鉴定，采用Reed-Muench法测定PDCoV的TCID₅₀，应用5×10^4.0 TCID₅₀/mL的病毒悬液接种11日龄仔猪并观察临床症状。结果显示，试验成功分离到1株PDCoV，将其命名为TJ₁株，该病毒的第5代病毒效价为10^4.5 TCID₅₀/mL；TJ₁株对脂溶剂氯仿和乙醚不敏感，耐酸，在pH 3.0的环境下稳定，对热较敏感，在50℃条件下1 h便可将其灭活；其M基因和GenBank中已发表的PDCoV M基因核苷酸序列同源性为94%~99%；致病力试验结果显示，该病毒可引起仔猪发生腹泻，发病率100%。本研究成功分离了1株PDCoV毒株TJ₁株，对该分离株生物学特性进行初步研究，为进一步开展PDCoV相关研究奠定了基础。

关键词: 猪德尔塔冠状病毒(PDCoV); 分离; 鉴定; 生物学特性

Abstract:

The aim of this study was to obtain porcine deltacoronavirus (PDCoV) strain in China and study its biological characteristics. Porcine kidney (PK-15) cell monolayers were inoculated with the samples that were positive for PDCoV only by RT-PCR. PDCoV from the first passage was further serially passed in PK-15 cells. These cell cultures infected with PDCoV were confirmed by cytopathic effect (CPE), sequencing and analysis of PDCoV M gene, acid test; TCID₅₀ was confirmed by Reed-Muench method; The 11-day-old piglets were inoculated with 5×10^4.0 TCID₅₀/mL virus suspension and observed for clinical symptoms. The results indicated that the PDCoV was successfully isolated in cell culture and designated TJ₁, and the infectious titer of the 5th passage of TJ₁ strain was 10^4.5 TCID₅₀/mL; The isolated virus was resistant to chloroform and ether, it was stable at pH 3.0, and the virus was inactivated at 50℃ for 1 h; The TJ₁ M gene shared a homology of 94% to 99% with PDCoV M gene sequences published previously in GenBank; The animal pathogenicity test showed that diarrhea occurred in all piglets by TJ₁. This study successfully isolated a PDCoV TJ₁ strain, and these biological characteristics of the strain were studied preliminarily, which provided a foundation for further research on PDCoV.

Key words: porcine deltacoronavirus (PDCoV); isolation; identification; biological characteristics

中图分类号:

S858.28

郑丽, 李秀丽, 鄢明华, 任卫科, 张蕾, 路超, 田向学, 韩伟. 猪德尔塔冠状病毒TJ₁株的分离鉴定及生物学特性分析[J]. 《中国畜牧兽医》, 2018, 45(1): 219-224.

ZHENG Li, LI Xiuli, YAN Minghua, REN Weike, ZHANG Lei, LU Chao, TIAN Xiangxue, HAN Wei. Isolation, Identification and Biological Characteristics Analysis of Porcine Deltacoronavirus TJ₁[J]. , 2018, 45(1): 219-224.

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猪德尔塔冠状病毒TJ₁株的分离鉴定及生物学特性分析

Isolation, Identification and Biological Characteristics Analysis of Porcine Deltacoronavirus TJ₁

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