重庆地区犬圆环病毒检测及序列分析

doi:10.16431/j.cnki.1671-7236.2020.10.004

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (10): 3069-3077.doi: 10.16431/j.cnki.1671-7236.2020.10.004

重庆地区犬圆环病毒检测及序列分析

闫修魁^1,2, 韩知晓^1,2, 杜倩^1,2, 汪伟^1,2,3, 朱远致^1,2, 辛佳亮^1,2, 易驰喆^1,2,4, 闭璟珊², 孙文超^3,5, 郑敏²

1. 广西大学动物科学技术学院, 南宁 530001;
2. 广西壮族自治区动物疫病预防控制中心, 南宁 530004;
3. 军事科学院军事兽医研究所, 长春 130122;
4. 柳州市工人医院, 柳州 545005;
5. 温州大学病毒学研究所, 温州 325035

收稿日期:2020-04-18 出版日期:2020-10-20 发布日期:2020-10-17
通讯作者: 孙文超, 郑敏 E-mail:sunwnechao131@163.com;zhgmn26@163.com
作者简介:闫修魁(1995-),男,重庆人,硕士生,研究方向:动物传染病学,E-mail:184500951@qq.com
基金资助:
广西壮族自治区自然科学基金项目（2012GXNSFAA053073）；浙江省自然科学基金青年基金项目（LQ19C180001）；广西农业科技自筹经费项目（Z201986）

Detection and Sequence Analysis of Canine Circovirus in Chongqing

YAN Xiukui^1,2, HAN Zhixiao^1,2, DU Qian^1,2, WANG Wei^1,2,3, ZHU Yuanzhi^1,2, XIN Jialiang^1,2, YI Chizhe^1,2,4, BI Jingshan², SUN Wenchao^3,5, ZHENG Min²

1. College of Animal Science and Technology, Guangxi University, Nanning 530001, China;
2. Guangxi Center for Animal Disease Control and Prevention, Nanning 530004, China;
3. Institute of Military Veterinary, The Academy of Military Medical Sciences, Changchun 130122, China;
4. Worker's Hospital of Liuzhou City, Liuzhou 545005, China;
5. Institute of Virology, Wenzhou University, Wenzhou 325035, China

Received:2020-04-18 Online:2020-10-20 Published:2020-10-17

摘要/Abstract

摘要： 本试验旨在调查近年来新发的犬圆环病毒（canine circovirus，CCV）在重庆地区的流行情况，探索重庆流行毒株的特性。本研究利用PCR方法对2017年采集于重庆地区的100份犬血清样品进行CCV核酸检测，对所得CCV阳性样品进行全基因组扩增与测序，并利用MegAlign、Mega 6.0和RDP4等软件对分离到的重庆CCV毒株进行分析。结果显示，重庆地区100份犬血清中共有4份为CCV阳性，阳性率为4%，从4份阳性样品中共获得3株不同的CCV基因组（CQ76、CQ79和CQ82），全长基因组序列均为2 062 nt，与此前报道长度为2 063 nt的CCV基因组相比缺少了一个碱基，该碱基位于5'-基因间隔区内茎环结构的下游。3个毒株的两个ORF之间5'末端间隔区和3'末端间隔区长度分别为134（1 929-2 062 nt）和203 nt（913-1 115 nt）。CQ76、CQ79和CQ82的同源性在99.8%~100%之间，其中CQ76与CQ82仅在第2 019位点存在同义突变；所获得的3个重庆毒株与国内外报道的其他CCV基因组序列同源性为82.7%~97.1%，其中，ORF2的变异程度大于ORF1。基于病毒ORF2序列和全基因组分别构建NJ进化树中，CQ76、CQ79和CQ82均属于同一亚群，与属于基因Ⅱ型的中国毒株204株遗传距离较近，同源性为96.5%~96.7%。RDP4重组分析发现，CQ76、CQ79和CQ82毒株基因组均为广西毒株204株和388株的重组序列，重组区域坐落于ORF2。本研究为丰富CCV流行病学和遗传进化信息，以及进一步防控研究提供基础数据。

关键词: 犬圆环病毒(CCV); 序列分析; 同源性分析

Abstract: The aim of this study was to investigate the prevalence of canine circovirus (CCV) in Chongqing in recent years,and to explore the characteristics of the field strains in Chongqing.In this study,100 dog serum samples collected in Chongqing in 2017 were tested for CCV nucleic acid by PCR,and the obtained CCV positive samples were amplified and sequenced with the full length genome,and the Chongqing CCV strain isolated was analyzed by MegAlign,Mega 6.0 and RDP4 software.Four positive sample were found,with a positive rate of 4%,and three full length CCV genomes (CQ76,CQ79,and CQ82) were obtained,with a total length of 2 062 nt.Compared with the previously reported genome of CCV with a length of 2 063 nt,one base was missing,which was located downstream of the stem ring structure in the 5'-intergenic region.The length of the 5'-intergenic region and 3'-intergenic region between the two ORFs of the three strains were 134 (1 929-2 062 nt) and 203 nt (913-1 115 nt),respectively.The homology of CQ76,CQ79,and CQ82 ranged from 99.8% to 100%,among which,CQ76 and CQ82 only had synonymous mutations at the 2 019 locus.The homology of genome sequences of the three Chongqing strains and other strains was 82.7% to 97.1%,among which,the degree of variation of ORF2 was greater than that of ORF1.Based on virus ORF2 sequences and genome-wide respectively build Neighbor-Joining in the evolutionary tree,CQ76,CQ79 and CQ82 all belonged to the same subgroup,which was close to the Chinese strain 204,which belonged to genotype Ⅱ,and the homology was 96.5% to 96.7%.RDP4 recombinant analysis indicated that the genomes of CQ76,CQ79 and CQ82 strains were all the recombinant sequences of 204 and 388 strains from Guangxi,and the recombinant region was located in ORF2.Above results enriched the epidemiological and genetic evolution information of CCV and provided further basic data for prevention and control research.

Key words: canine circovirus (CCV); sequence analysis; homology analysis

中图分类号:

S852.65

闫修魁, 韩知晓, 杜倩, 汪伟, 朱远致, 辛佳亮, 易驰喆, 闭璟珊, 孙文超, 郑敏. 重庆地区犬圆环病毒检测及序列分析[J]. 中国畜牧兽医, 2020, 47(10): 3069-3077.

YAN Xiukui, HAN Zhixiao, DU Qian, WANG Wei, ZHU Yuanzhi, XIN Jialiang, YI Chizhe, BI Jingshan, SUN Wenchao, ZHENG Min. Detection and Sequence Analysis of Canine Circovirus in Chongqing[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3069-3077.

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重庆地区犬圆环病毒检测及序列分析

Detection and Sequence Analysis of Canine Circovirus in Chongqing

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