2013～2017年江西省猪圆环病毒2型分子流行病学调查
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摘要
为掌握江西地区猪圆环病毒2型(PCV2)的分子流行病学及其遗传变异情况,本研究运用实验室已建立的PCR方法对2013～2017年采集自江西省南昌市、宜春市、新余市、赣州市、吉安市、九江市、上饶市、抚州市、景德镇市、鹰潭市等10个地区的1 082份疑似PCV2感染猪病料组织进行病原学检测,并通过PCR扩增、克隆和基因测序对PCV2的全基因组序列进行分析。研究表明,1 082份病料中有610份为PCV2阳性,总阳性率为56.4%,其中2013～2017年阳性率分别为52.7%、48.8%、58.5%、71.0%和67.4%。共获得89株PCV2全基因组序列,其中有83株为PCV2b基因型,其中53株归类于PCV2b-1C基因亚群,30株归为PCV2b-1A/1B基因亚群;6株为PCV2a型。测序毒株与参考毒株ORF2基因核苷酸、氨基酸序列同源性分别为88.9%～100.0%和86.0%～100.0%;氨基酸序列分析表明,Cap蛋白存在20个氨基酸突变位点。本研究表明,江西地区猪群中PCV2的主导基因型为PCV2b,其中又以PCV2b-1C基因亚群占据主导地位,同时也存在少量PCV2a基因型。
To investigate the molecular epidemiology and genetic diversity of porcine circovirus type 2(PCV2) in Jiangxi province,1 082 tissue samples were collected during 2013 to 2017 from suspected pigs in 10 regions(including Nanchang,Yichun,Xinyu,Ganjiang,Ji'an,Jiujiang,Shangrao,Fuzhou,Jingdezhen and Yingtan cities) of Jiangxi province were detected for PCV2.And PCV2 isolates complete genome sequences were amplified,cloned and sequenced.The results showed that 610 of 1 082 samples were detected to be positive of PCV2 and the total positive rate was 56.4%.The positive rates from 2013 to 2017 were 52.7%,48.8%,58.5%,71.0% and 67.4%,respectively.The 83 of 89 PCV2 isolates belonged to PCV2 b,and 53 and 30 of 83 isolates were grouped into PCV2 b-1 C and PCV2 b-1 A/1 B,respectively,and 6 isolates belonged to PCV2 a.The homology of nucleotides and the deduced amino acid sequence of ORF2 between sequenced strains and reference strains were 88.9% to 100.0% and 86.0% to 100.0%,respectively.Analysis of Cap acid amino sequences showed that there were 20 amino acid mutation sites.This research indicated that the dominant genotype of PCV2 in pigs in Jiangxi was PCV2 b,and PCV2 b-1 C was a dominant subgenotype.At the same time,a small amount of PCV2 a subgenotype also existed.
To investigate the molecular epidemiology and genetic diversity of porcine circovirus type 2(PCV2) in Jiangxi province,1 082 tissue samples were collected during 2013 to 2017 from suspected pigs in 10 regions(including Nanchang,Yichun,Xinyu,Ganjiang,Ji'an,Jiujiang,Shangrao,Fuzhou,Jingdezhen and Yingtan cities) of Jiangxi province were detected for PCV2.And PCV2 isolates complete genome sequences were amplified,cloned and sequenced.The results showed that 610 of 1 082 samples were detected to be positive of PCV2 and the total positive rate was 56.4%.The positive rates from 2013 to 2017 were 52.7%,48.8%,58.5%,71.0% and 67.4%,respectively.The 83 of 89 PCV2 isolates belonged to PCV2 b,and 53 and 30 of 83 isolates were grouped into PCV2 b-1 C and PCV2 b-1 A/1 B,respectively,and 6 isolates belonged to PCV2 a.The homology of nucleotides and the deduced amino acid sequence of ORF2 between sequenced strains and reference strains were 88.9% to 100.0% and 86.0% to 100.0%,respectively.Analysis of Cap acid amino sequences showed that there were 20 amino acid mutation sites.This research indicated that the dominant genotype of PCV2 in pigs in Jiangxi was PCV2 b,and PCV2 b-1 C was a dominant subgenotype.At the same time,a small amount of PCV2 a subgenotype also existed.
引文
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[2] CHAE C.An emerging porcine circovirus type 2b mutant (mPCV2b) originally known as PCV2d[J].Veterinary Journal,2015,203(1):6-9.
[3] NOVOSEL D,TUBOLY T,CSAGOLA A,et al.Origin of porcine circovirus type 2 (PCV2) from swine affected by PCV2-associated diseases in Croatia[J].Veterinary Record,2014,174(17):431.
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[7] FRANZO G,CORTEY M,SEGALES J,et al.Phylodynamic analysis of porcine circovirus type 2:Methodological approach and datasets[J].Data in Brief,2016,8:549-552.
[8] OLVERA A,CORTEY M,SEGALES J.Molecular evolution of porcine circovirus type 2 genomes:Phylogeny and clonality[J].Virology,2007,357(2):175-185.
[9] 李玲,李国新,周艳君,等.2008—2011年中国部分地区猪圆环病毒2型的分子流行病学调查[J].中国动物传染病学报,2012,20(2):1-10. LI L,LI G X,ZHOU Y J,et al.Molecular epidemiological investigation into porcine circovirus type 2 between 2008 and 2011 in some regions of China[J].Chinese Journal of Animal Infectious Diseases,2012,20(2):1-10.(in Chinese)
[10] AN D J,ROH I S,SONG D S,et al.Phylogenetic characterization of porcine circovirus type 2 in PMWS and PDNS Korean pigs between 1999 and 2006[J].Veterinary Research,2007,129(2):115-122.
[11] FRANZO G,CORTEY M,OLVERA A,et al.Revisiting the taxonomical classification of porcine circovirus type 2 (PCV2):Still a real challenge[J].Virology Journal,2015,12:131.
[12] 吴瑷,章红兵,王鑫炎,等.2007～2012年浙江省及周边地区猪圆环病毒2型(PCV2)分子流行病学调查[J].农业生物技术学报,2013,21(4):456-463. WU Y,ZHANG H B,WANG X Y,et al.Molecular epidemiological investigation of porcine circovirus type 2(PCV2) in Zhejiang province and surrounding areas[J].Journal of Agricultural Biotechnology,2013,21(4):456-463.(in Chinese)
[13] HARMON K M,GAUGER P C,ZHANG J,et al.Whole-genome sequences of novel porcine circovirus type 2 viruses detected in swine from Mexico and the United States[J].Genome Announcement,2015,3(6):1315.
[14] GILLESPIE J,OPRIESSNIG T,MENG X J,et al.Porcine circovirus type 2 and porcine circovirus-associated disease[J].Journal of Veterinary Internal Medicine,2009,23(6):1151-1163.
[15] 廖启顺,李志祥,丁联成,等.2009-2010年部分猪场主要传染病流行病学调查[J].中国畜牧兽医,2012,39(2):194-199. LIAO Q S,LI Z X,DING L C,et al.Epidemiological research of major infectious disease from partial pig farm from 2009 to 2010[J].China Animal Husbandry & Veterinary Medicine,2012,39(2):194-199.(in Chinese)
[16] 邵萌,吕亚辉,杜谦,等.2010—2012年陕西地区猪圆环病毒2型分子流行病学调查[J].西北农林科技大学学报(自然科学版),2013,41(10):19-26. SHAO M,LYU Y H,DU Q,et al.Molecular epidemiological investigation on porcine circovirus type 2 in Shaanxi province during 2010-2012[J].Journal of Northwest A & F University (Natural Science Edition),2013,41(10):19-26.(in Chinese)
[17] SEGALES J,OLVERA A,GRAU-ROMA L,et al.PCV-2 genotype definition and nomenclature[J].Veterinary Record,2008,162(26):867-868.
[18] 王利丽,郑丽,路超,等.猪圆环病毒2型天津及周边地区分离株的遗传演化分析[J].中国畜牧兽医,2018,45(5):1331-1340. WANG L L,ZHENG L,LU C,et al.Phylogenetic analysis of porcine circovirus type 2 isolate from Tianjin and its surrounding areas[J].China Animal Husbandry & Veterinary Medicine,2018,45(5):1331-1340.(in Chinese)
[19] 戴爱玲,刘建奎,李鸿文,等.闽西地区猪圆环病毒2型血清学调查及遗传变异分析[J].中国兽医学报,2016,36(6):896-901. DAI A L,LIU J K,LI H W,et al.Serological investigation and genetic diversity of porcine circovirus type 2 infection in swine farms of Western Fujian pro-vince[J].Chinese Journal of Veterinary Science,2016,36(6):896-901.(in Chinese)
[20] 时庆贺,石昂,李新果,等.河南省2016-2017年致病性猪圆环病毒遗传演化分析[J].中国兽医学报,2018,38(5):857-862. SHI Q H,SHI A,LI X G,et al.Phylogenetic analysis of prevailing pathogenic porcine circovirus in Henan province during 2016-2017[J].Chinese Journal of Veterinary Science,2018,38(5):857-862.(in Chinese)
[21] 徐朋丽,张宇,韩昊莹,等.猪圆环病毒2型河南株全基因组的克隆与遗传进化分析[J].中国预防兽医学报,2016,38(12):989-992. XU P L,ZHANG Y,HAN H Y,et al.Cloning and phylogenetic evolution analysis of the complete genomes of porcine circovirus type 2 from Henan province[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(12):989-992.(in Chinese)
[22] KWEON C H,NGUYEN L T,YOO M S,et al.Differential recognition of the ORF2 region in a complete genome sequence of porcine circovirus type 2 (PCV2) isolated from boarbone marrow in Korea[J].Gene,2015,569(2):308-312.
[23] 赵军,周桐枫,刘鹏娟,等.四川地区腹泻仔猪群猪圆环病毒Ⅱ型分子流行病学调查[J].中国预防兽医学报,2016,38(7):546-549. ZHAO J,ZHOU T F,LIU P J,et al.Molecular epidemiology of porcine circovirus typeⅡ in diarrhea piglets in Sichuan region[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(7):546-549.(in Chinese)
[2] CHAE C.An emerging porcine circovirus type 2b mutant (mPCV2b) originally known as PCV2d[J].Veterinary Journal,2015,203(1):6-9.
[3] NOVOSEL D,TUBOLY T,CSAGOLA A,et al.Origin of porcine circovirus type 2 (PCV2) from swine affected by PCV2-associated diseases in Croatia[J].Veterinary Record,2014,174(17):431.
[4] KOINIG H C,TALKER S C,STADLER M,et al.PCV2 vaccination induces IFN-γ/TNF-α co-producing T cells with a potential role in protection[J].Veterinary Research,2015,46:20-24.
[5] 邓静,赵春容,曹三杰,等.2010—2013年四川地区猪圆环病毒2型的分子流行病学调查[J].中国兽医学报,2014,34(12):1868-1876. DENG J,ZHAO C R,CAO S J,et al.Molecular epidemiological investigate of porcine circovirus type 2 between 2010 and 2013 in some regions of Sichuan[J].Chinese Journal of Veterinary Science,2014,34(12):1868-1876.(in Chinese)
[6] TISCHER I,BODE L,PETERS D,et al.Distribution of antibodies to porcine circovirus in swine populations of different breeding farms[J].Archives of Virology,1995,140(4):737-743.
[7] FRANZO G,CORTEY M,SEGALES J,et al.Phylodynamic analysis of porcine circovirus type 2:Methodological approach and datasets[J].Data in Brief,2016,8:549-552.
[8] OLVERA A,CORTEY M,SEGALES J.Molecular evolution of porcine circovirus type 2 genomes:Phylogeny and clonality[J].Virology,2007,357(2):175-185.
[9] 李玲,李国新,周艳君,等.2008—2011年中国部分地区猪圆环病毒2型的分子流行病学调查[J].中国动物传染病学报,2012,20(2):1-10. LI L,LI G X,ZHOU Y J,et al.Molecular epidemiological investigation into porcine circovirus type 2 between 2008 and 2011 in some regions of China[J].Chinese Journal of Animal Infectious Diseases,2012,20(2):1-10.(in Chinese)
[10] AN D J,ROH I S,SONG D S,et al.Phylogenetic characterization of porcine circovirus type 2 in PMWS and PDNS Korean pigs between 1999 and 2006[J].Veterinary Research,2007,129(2):115-122.
[11] FRANZO G,CORTEY M,OLVERA A,et al.Revisiting the taxonomical classification of porcine circovirus type 2 (PCV2):Still a real challenge[J].Virology Journal,2015,12:131.
[12] 吴瑷,章红兵,王鑫炎,等.2007～2012年浙江省及周边地区猪圆环病毒2型(PCV2)分子流行病学调查[J].农业生物技术学报,2013,21(4):456-463. WU Y,ZHANG H B,WANG X Y,et al.Molecular epidemiological investigation of porcine circovirus type 2(PCV2) in Zhejiang province and surrounding areas[J].Journal of Agricultural Biotechnology,2013,21(4):456-463.(in Chinese)
[13] HARMON K M,GAUGER P C,ZHANG J,et al.Whole-genome sequences of novel porcine circovirus type 2 viruses detected in swine from Mexico and the United States[J].Genome Announcement,2015,3(6):1315.
[14] GILLESPIE J,OPRIESSNIG T,MENG X J,et al.Porcine circovirus type 2 and porcine circovirus-associated disease[J].Journal of Veterinary Internal Medicine,2009,23(6):1151-1163.
[15] 廖启顺,李志祥,丁联成,等.2009-2010年部分猪场主要传染病流行病学调查[J].中国畜牧兽医,2012,39(2):194-199. LIAO Q S,LI Z X,DING L C,et al.Epidemiological research of major infectious disease from partial pig farm from 2009 to 2010[J].China Animal Husbandry & Veterinary Medicine,2012,39(2):194-199.(in Chinese)
[16] 邵萌,吕亚辉,杜谦,等.2010—2012年陕西地区猪圆环病毒2型分子流行病学调查[J].西北农林科技大学学报(自然科学版),2013,41(10):19-26. SHAO M,LYU Y H,DU Q,et al.Molecular epidemiological investigation on porcine circovirus type 2 in Shaanxi province during 2010-2012[J].Journal of Northwest A & F University (Natural Science Edition),2013,41(10):19-26.(in Chinese)
[17] SEGALES J,OLVERA A,GRAU-ROMA L,et al.PCV-2 genotype definition and nomenclature[J].Veterinary Record,2008,162(26):867-868.
[18] 王利丽,郑丽,路超,等.猪圆环病毒2型天津及周边地区分离株的遗传演化分析[J].中国畜牧兽医,2018,45(5):1331-1340. WANG L L,ZHENG L,LU C,et al.Phylogenetic analysis of porcine circovirus type 2 isolate from Tianjin and its surrounding areas[J].China Animal Husbandry & Veterinary Medicine,2018,45(5):1331-1340.(in Chinese)
[19] 戴爱玲,刘建奎,李鸿文,等.闽西地区猪圆环病毒2型血清学调查及遗传变异分析[J].中国兽医学报,2016,36(6):896-901. DAI A L,LIU J K,LI H W,et al.Serological investigation and genetic diversity of porcine circovirus type 2 infection in swine farms of Western Fujian pro-vince[J].Chinese Journal of Veterinary Science,2016,36(6):896-901.(in Chinese)
[20] 时庆贺,石昂,李新果,等.河南省2016-2017年致病性猪圆环病毒遗传演化分析[J].中国兽医学报,2018,38(5):857-862. SHI Q H,SHI A,LI X G,et al.Phylogenetic analysis of prevailing pathogenic porcine circovirus in Henan province during 2016-2017[J].Chinese Journal of Veterinary Science,2018,38(5):857-862.(in Chinese)
[21] 徐朋丽,张宇,韩昊莹,等.猪圆环病毒2型河南株全基因组的克隆与遗传进化分析[J].中国预防兽医学报,2016,38(12):989-992. XU P L,ZHANG Y,HAN H Y,et al.Cloning and phylogenetic evolution analysis of the complete genomes of porcine circovirus type 2 from Henan province[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(12):989-992.(in Chinese)
[22] KWEON C H,NGUYEN L T,YOO M S,et al.Differential recognition of the ORF2 region in a complete genome sequence of porcine circovirus type 2 (PCV2) isolated from boarbone marrow in Korea[J].Gene,2015,569(2):308-312.
[23] 赵军,周桐枫,刘鹏娟,等.四川地区腹泻仔猪群猪圆环病毒Ⅱ型分子流行病学调查[J].中国预防兽医学报,2016,38(7):546-549. ZHAO J,ZHOU T F,LIU P J,et al.Molecular epidemiology of porcine circovirus typeⅡ in diarrhea piglets in Sichuan region[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(7):546-549.(in Chinese)
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