猪德尔塔冠状病毒CH7328株的致病性及全基因组序列分析

doi:10.16431/j.cnki.1671-7236.2021.11.024

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (11): 4143-4153.doi: 10.16431/j.cnki.1671-7236.2021.11.024

猪德尔塔冠状病毒CH7328株的致病性及全基因组序列分析

王征帆^1,2,3, 朱利塞^1,3, 李菲^1,3, 向蕊^1,2,3, 李祎云^1,2,3, 应碧云^1,2,3, 王贵平^1,3,4, 白挨泉², 贾爱卿^1,3,4

1. 广东海大畜牧兽医研究院, 广州 511400;
2. 佛山科学技术学院, 佛山 528231;
3. 广东省养猪与猪病防控技术研究企业重点实验室, 广州 511400;
4. 广东海大集团股份有限公司, 广州 511400

收稿日期:2021-04-23 出版日期:2021-11-20 发布日期:2021-11-01
通讯作者: 白挨泉, 贾爱卿 E-mail:baiaiquan2008@163.com;59492815@qq.com
作者简介:王征帆(1993-),男,贵州雷山人,硕士,研究方向:猪传染病诊断与防治,E-mail:15320520429@163.com
基金资助:
广东省重点领域研发计划项目（2019B020218004）；韶关市生猪优势产区产业园；广州市科学研究计划重点项目（201804020006）；番禺区创新创业领军团队项目（2017-R02-4）；广东省科技计划项目（2020B1212070023）；猪用疫苗生产新工艺及产业化关键技术的研发项目（2017YFD0500604）；广东省教育厅预防兽医学重点实验室项目（2014KTSPT037）

Pathogenicity and Whole Genome Sequence Analysis of Porcine Deltacoronavirus CH7328 Strain

WANG Zhengfan^1,2,3, ZHU Lisai^1,3, LI Fei^1,3, XIANG Rui^1,2,3, LI Yiyun^1,2,3, YING Biyun^1,2,3, WANG Guiping^1,3,4, BAI Aiquan², JIA Aiqing^1,3,4

1. Guangdong Haida Institute of Animal Husbandry & Veterinary, Guangzhou 511400, China;
2. Foshan University, Foshan 528231, China;
3. Guangdong Provincial Key Laboratory of Pig Breeding and Pig Disease Prevention and Control Technology Research Enterprise, Guangzhou 511400, China;
4. Guangdong Haid Group Co., Ltd., Guangzhou 511400, China

Received:2021-04-23 Online:2021-11-20 Published:2021-11-01

摘要/Abstract

摘要： 为了解猪德尔塔冠状病毒（Porcine deltacoronavirus，PDCoV） CH7328株的致病性，并分析其基因变异和遗传演变规律，试验通过病毒培养、间接免疫荧光试验（IFA）、病毒生长曲线及仔猪致病性试验鉴定病毒的生长特性和致病性，并分析该毒株的全基因组序列。结果显示，感染PDCoV CH7328株的ST细胞出现肿大变圆、细胞呈碎玻璃状且拉丝、细胞集聚成簇和脱落等病变。IFA结果显示，PDCoV CH7328株主要分布在细胞质中。生长曲线结果显示，PDCoV CH7328株在6 h就已开始增殖，在36 h时达到峰值，对应的TCID₅₀为10^-8.0/mL，随后逐渐下降。仔猪致病性结果显示，攻毒组仔猪出现腹泻、脱水和厌食等临床症状，仔猪小肠明显变薄、透明而充盈，小肠绒毛严重脱落、萎缩或减少、肠绒毛细胞坏死等。粪便病原检测结果显示，在攻毒第2天便能检测到病毒，之后排毒持续增加，而正常对照仔猪无任何临床现象。全基因组测序及序列分析显示，PDCoV CH7328株的基因组全长为25 420 bp，与国内GD株的相似性最高，达到99.9%；遗传演化结果显示，PDCoV CH7328株与国内HKU15-155、CHN-HN-2014和GD株处在同一进化分支，亲缘关系最近。本研究结果为中国PDCoV疫苗候选株的筛选及猪场疫病防控提供了参考数据和理论依据。

关键词: 猪德尔塔冠状病毒(PDCoV); CH7328株; 致病性; 全基因序列分析

Abstract: In order to understand the pathogenicity of Porcine deltacoronavirus (PDCoV) CH7328 strain, and analyze its genetic variation and genetic evolution, this test used virus culture, indirect immunofluorescence assay (IFA), virus growth curve and piglet pathogenicity test to identify the growth characteristics and pathogenicity of the virus, and analyze the entire genome sequence of the virus strain. The results showed that ST cells infected with PDCoV CH7328 strain showed swelling and rounding, the cells were broken glass-like and brushed, and the cells clustered and fell off. The results of the IFA test showed that the PDCoV CH7328 strain was mainly distributed in the cytoplasm. The growth curve results showed that the PDCoV CH7328 strain began to proliferate at 6 h, and reached a peak at 36 h, the corresponding TCID₅₀ was 10^-8.0/mL, and then gradually decreased. The pathogenicity results of piglets showed that the piglets in challenge group showed clinical symptoms such as diarrhea, dehydration and anorexia, the small intestine was obviously thinner, transparent and full, and the small intestinal villi were severely shed, atrophy or reduced, and intestinal villi cells were necrotic. The results of fecal pathogen detection showed that the virus could be detected on the second day of the challenge, after which the detoxification continued to increase, while the normal control piglets did not have any clinical signs. Whole genome sequencing and sequence analysis showed that the full length of the genome of the PDCoV CH7328 strain was 25 420 bp, which was the highest similar to the domestic GD strain, reaching 99.9%. The genetic evolution results showed that the PDCoV CH7328 strain was in the same evolutionary branch with domestic HKU15-155, CHN-HN-2014 and GD strains, and the genetic relationship was the closest. The results of this study provided a reference data and theoretical basis for the screening of PDCoV vaccine candidate strains and the prevention and control of pig farm diseases in my country.

Key words: Porcine deltacoronavirus (PDCoV); CH7328 strain; pathogenicity; whole gene sequence analysis

中图分类号:

S852.65⁺1

王征帆, 朱利塞, 李菲, 向蕊, 李祎云, 应碧云, 王贵平, 白挨泉, 贾爱卿. 猪德尔塔冠状病毒CH7328株的致病性及全基因组序列分析[J]. 中国畜牧兽医, 2021, 48(11): 4143-4153.

WANG Zhengfan, ZHU Lisai, LI Fei, XIANG Rui, LI Yiyun, YING Biyun, WANG Guiping, BAI Aiquan, JIA Aiqing. Pathogenicity and Whole Genome Sequence Analysis of Porcine Deltacoronavirus CH7328 Strain[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4143-4153.

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猪德尔塔冠状病毒CH7328株的致病性及全基因组序列分析

Pathogenicity and Whole Genome Sequence Analysis of Porcine Deltacoronavirus CH7328 Strain

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