利用宏病毒组学对鹅痛风病原的鉴定

doi:10.16431/j.cnki.1671-7236.2021.12.034

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (12): 4661-4670.doi: 10.16431/j.cnki.1671-7236.2021.12.034

利用宏病毒组学对鹅痛风病原的鉴定

王安平¹, 郝福星¹, 张硕², 谢军¹, 刘莉¹, 朱善元¹

1. 江苏农牧科技职业学院, 江苏省兽用生物制药高技术研究重点实验室, 泰州 225300;
2. 江苏海洋大学食品科学与工程学院, 连云港 222005

收稿日期:2021-06-08 出版日期:2021-12-20 发布日期:2021-12-02
通讯作者: 朱善元 E-mail:wap4017@163.com
作者简介:王安平(1980-),女,江苏泰州人,博士,教授,研究方向:预防兽医,E-mail:704540927@qq.com;郝福星(1982-),男,江苏徐州人,副教授,研究方向:基础兽医,E-mail:vethfx@163.com
基金资助:
江苏省农业科技自主创新项目（CX（18）1004）；江苏省现代农业产业技术体系集成创新中心（JATS[2020]345）；江苏高校"青蓝工程"（苏教师函[2020]10号）；江苏省高等学校自然科学研究重大项目（18KJA230001）

Identification of Pathogen of Gosling Gout Using Viral Metagenomics

WANG Anping¹, HAO Fuxing¹, ZHANG Shuo², XIE Jun¹, LIU Li¹, ZHU Shanyuan¹

1. Jiangsu Key Laboratory of Veterinary Bio-Pharmaceutical High Technology Research, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China;
2. School of Food Science and Engineering, Jiangsu Ocean University, Lianyungang 222005, China

Received:2021-06-08 Online:2021-12-20 Published:2021-12-02

摘要/Abstract

摘要： 为确定2020年江苏盐城一鹅场雏鹅痛风的病原,试验对病料样品进行宏病毒组测序,通过比对分析原始数据发现疑似病原;通过从头组装获得其全基因组序列,并进行相似性比对和遗传进化分析;通过对疑似病毒的分离、鉴定和动物回归试验进行验证。宏病毒组学测序结果显示,在获得的全部599 733条原始读长(reads)中,病毒reads有278 593条,其中鹅星状病毒序列占95%,未发现其他疑似病毒序列;经从头组装获得了一条完整的鹅星状病毒全基因组序列,命名为YC20,该病毒基因组全长7 137 nt,GenBank登录号为:MW536497,基因组结构及特征与其他星状病毒相同。全基因组序列相似性比对结果显示,YC20与新型鹅星状病毒SD01相似性最高,达98.2%;遗传进化分析结果显示,YC20与其他新型鹅星状病毒在同一分支,属于新型鹅星状病毒。禽胚分离结果显示,用鹅胚进行病原分离时,前3代次未出现死亡,第5代次后,可对鹅胚100%致死,而用SPF鸡胚或SPF鸭胚分离病原时,均未出现病变或死亡,PCR检测亦为阴性;纯化的尿囊液经负染后,电镜下可见大小约25 nm的病毒粒子。雏鹅回归试验结果显示,YC20接种3日龄雏鹅后,第6~8天出现死亡高峰期,死亡率为58.3%,剖检死亡鹅可见心脏、肝脏表面覆盖一层白色包膜,膝关节有大量白色颗粒样沉淀,肾脏充血肿大。以上结果说明,该鹅场痛风的病原为新型鹅星状病毒。本研究将宏病毒组学与传统的病毒分离鉴定技术相结合,证实了鹅痛风的病原,对病原的鉴定和研究具有指导意义。

关键词: 痛风; 鹅星状病毒; 宏病毒组学; 分离; 鉴定

Abstract: In order to determine the causative agent of gosling gout in a goose farm in Yancheng city in 2020, the samples were deeply sequenced by the method of viral metagenomics, and the suspected pathogens were found through the alignment of the original data.The whole genome sequence was obtained by de novo assembly, and the homology and phylogenetic analysis were done.The pathogen was verified by the isolation and identification of the suspected virus and animal regression test.The results of viral metagenome sequencing showed that a total of 599 733 original reads were obtained.There were 278 593 virus reads, of which Goose astrovirus sequences accounted for 95%, no other suspected virus sequences were found.The whole genome sequence was obtained by de novo assembly, which was 7 137 nt in length with the typical genomic characterization of other known avastroviruses, the isolated virus was named YC20, GenBank accession No.:MW536497.The results of similarity alignment showed that the YC20 isolate shared the highest identity of 98.2% with the Goose astrovirus strain SD01 in terms of the whole-genome sequence.The results of phylogenetic analysis showed that YC20 clustered together with the new type of Goose astrovirus.The results of virus isolation showed that no goose embryos died during their first 3 passages but the mortality rate reached 100% after the fifth passages.Meanwhile, no death or pathological changes were found in SPF chicken or duck embryos, and PCR results were negative.After negative staining, the virus particles of about 25 nm were observed under electron microscope.In the goose regression experiment, the infected goslings experienced a significant peak of death during days 6 to 8, resulting in a mortality rate of 58.3%.At necropsy, uric deposits in heart, liver, and articular cavity, as well as severe hemorrhage and kidney swelling were observed in dead goslings.The results suggested that the pathogen of gout in this goose farm was a new type of the new Goose astrovirus.In this study, the combination of viral metagenomics with traditional virus isolation techniques confirms the pathogen of goose gout, which had guiding significance for the identification and study of the pathogen.

Key words: gout; Goose astrovirus; viral metagenomics; isolation; identification

中图分类号:

S852.65⁺7

王安平, 郝福星, 张硕, 谢军, 刘莉, 朱善元. 利用宏病毒组学对鹅痛风病原的鉴定[J]. 中国畜牧兽医, 2021, 48(12): 4661-4670.

WANG Anping, HAO Fuxing, ZHANG Shuo, XIE Jun, LIU Li, ZHU Shanyuan. Identification of Pathogen of Gosling Gout Using Viral Metagenomics[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4661-4670.

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利用宏病毒组学对鹅痛风病原的鉴定

Identification of Pathogen of Gosling Gout Using Viral Metagenomics

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