猪A群轮状病毒SCJY-13株的分离鉴定及致病性分析

doi:10.16431/j.cnki.1671-7236.2022.02.028

中国畜牧兽医 ›› 2022, Vol. 49 ›› Issue (2): 660-668.doi: 10.16431/j.cnki.1671-7236.2022.02.028

猪A群轮状病毒SCJY-13株的分离鉴定及致病性分析

陈小飞¹, 张斌², 张春红³, 张建峰³, 廖明^1,3,4

1. 华南农业大学兽医学院, 人兽共患病防控制剂国家地方联合工程实验室, 广东省动物原性人兽共患病防控重点实验室, 农业农村部人兽共患病重点实验室, 广州 510642;
2. 西南民族大学生命科学与技术学院, 成都 610041;
3. 广东省农业科学院动物卫生研究所, 广州 510640;
4. 岭南现代农业科学与技术广东省实验室茂名分中心, 茂名 525000

收稿日期:2021-08-11 出版日期:2022-02-05 发布日期:2022-01-27
通讯作者: 廖明 E-mail:mliao@scau.edu.cn
作者简介:陈小飞,E-mail:chenxf-1@163.com
基金资助:
广东省重点领域研发计划项目(2020B0202080004);2020年度广东省农业科学院"杰出人才"培养项目(R2020PY-JC001)

Isolation, Identification and Pathogenicity Analysis of Porcine Rotavirus Group A Strain SCJY-13

CHEN Xiaofei¹, ZHANG Bin², ZHANG Chunhong³, ZHANG Jianfeng³, LIAO Ming^1,3,4

1. National and Regional Joint Engineering Laboratory for Medicament for Zoonoses Prevention and Control, Key Laboratory for Prevention and Control of Zoonotic Diseases of Guangdong Province, Key Laboratory of Zoonoses Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
2. College of Life Science and Technology, Southwest Minzu University, Chengdu 610041, China;
3. Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
4. Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming 525000, China

Received:2021-08-11 Online:2022-02-05 Published:2022-01-27

摘要/Abstract

摘要： 【目的】在从腹泻仔猪粪便中分离鉴定猪A群轮状病毒(Rotavirus group A, RVA)并了解其致病性。【方法】应用MA-104细胞从仔猪腹泻粪便中分离鉴定RVA, 将其经口感染健康初生仔猪, 观察其临床症状, 并利用实时荧光定量PCR检测排毒、HE染色观察病理变化、免疫组织化学(immunohistochemistry, IHC)试验了解病毒分布。【结果】分离到1株可引起MA-104细胞明显细胞病变的G9P[23] RVA, 命名为RVA/Pig-tc/CHN/SCJY-13/2017/G9P[23](简称SCJY-13株), 病毒滴度为10^5.5 TCID₅₀/100 μL。经口感染SCJY-13株的仔猪在感染后11 h出现腹泻, 持续到感染后165 h完全恢复, 其发病率为100%(7/7), 病死率为28.57%(2/7)。感染后8 h可从仔猪肛拭子检测到病毒核酸, 直至感染后192 h, 峰值出现在感染后24 h。感染后54 h各段小肠病毒载量达到最高, 其中回肠中病毒载量最高, 显著高于十二指肠、空肠(P<0.05);HE染色和IHC检测结果显示, SCJY-13在感染仔猪小肠的绒毛及隐窝中大量聚集, 尤其是回肠段; SCJY-13株感染引起十二指肠、空肠和回肠绒毛固有层大量淋巴细胞浸润、黏膜上皮细胞和肠绒毛尖端空泡变性、柱状细胞增多、绒毛断裂脱落等, 以回肠段较严重。【结论】 SCJY-13株能引起新生仔猪100%发病, 其主要靶位区在回肠, 感染后排毒时间长。本试验结果为猪RVA的致病性研究提供了一定的参考依据。

关键词: 猪; A群轮状病毒(RVA); G9P[23]; 分离鉴定; 致病性

Abstract: 【Objective】 This study was aimed to isolate and identify porcine Rotavirus group A (RVA) from feces of piglets suffered from diarrhea, and understand the pathogenicity of the isolate strain.【Method】 RVA was isolated and identified from piglet diarrhea feces by MA-104 cells, and orally infected healthy newborn piglets.The clinical symptoms were observed, the detoxification was detected by Real-time quantitative PCR, the pathological changes were observed by HE staining, and the distribution of virus was tested by immunohistochemistry (IHC).【Result】 A strain of G9P[23] RVA, named RVA/Pig-tc/CHN/SCJY-13/2017/G9P[23] (SCJY-13 strain for short), which could cause obvious cytopathy of MA-104 cells, was isolated, and the virus titer was 10^5.5 TCID₅₀/100 μL.Diarrhea occurred at 11 h post infection and lasted until 165 h post infection.The incidence rate was 100% (7/7), and the mortality rate was 28.57% (2/7).Viral nucleic acid could be detected from piglet anal swabs at 8 h post infection until 192 h post infection, and the peak appeared at 24 h post infection.At the same time, the viral load in each segment of small intestine reached the highest 54 h post infection, and the viral load in ileum was the highest, which was significantly higher than that in duodenum, jejunum and feces (P < 0.05).HE staining and IHC detection showed that SCJY-13 accumulated in villi and crypts of small intestine of infected piglets, especially in ileum.SCJY-13 strain infection caused a large number of lymphocyte infiltration in duodenum, jejunum and ileum villus lamina propria, mucosal epithelial cells and vacuolar degeneration at the tip of intestinal villus, increase of columnar cells, villus breaking and falling off, especially in ileum.【Conclusion】 The SCJY-13 strain could cause 100% morbidity of neonatal piglets and shed virus for a long time, and the virus was mainly distributed in the ileum of small intestine.These results provided a valuable reference for the pathogenicity study of procine RVA.

Key words: porcine; Rotavirus group A (RVA); G9P[23]; isolation and identification; pathogenicity

中图分类号:

S852.65⁺1

陈小飞, 张斌, 张春红, 张建峰, 廖明. 猪A群轮状病毒SCJY-13株的分离鉴定及致病性分析[J]. 中国畜牧兽医, 2022, 49(2): 660-668.

CHEN Xiaofei, ZHANG Bin, ZHANG Chunhong, ZHANG Jianfeng, LIAO Ming. Isolation, Identification and Pathogenicity Analysis of Porcine Rotavirus Group A Strain SCJY-13[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 660-668.

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猪A群轮状病毒SCJY-13株的分离鉴定及致病性分析

Isolation, Identification and Pathogenicity Analysis of Porcine Rotavirus Group A Strain SCJY-13

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