2017年江西部分地区PEDV分子流行病学调查

doi:10.16431/j.cnki.1671-7236.2019.05.026

摘要/Abstract

摘要：

为了解江西地区猪流行性腹泻病毒（PEDV）的流行和变异情况，本研究利用RT-PCR方法对2017年采自江西省部分地区规模化猪场182份腹泻仔猪小肠组织和粪便样品进行检测，并设计了2对引物对37份阳性病料的S基因进行扩增、克隆及序列测定，以及与GenBank中登录的22株PEDV S基因参考序列进行遗传进化分析。结果显示，37株PEDV江西流行株的S基因序列长为4 158或4 161 bp，编码1 385或1 386个氨基酸，全部为Group 1型，与美国流行毒株较为接近，而与欧洲毒株（Br1/87）及疫苗株（CV777）亲缘关系较远；37株PEDV中有36株为G1-1亚群，1株为G1-2亚群；37株PEDV江西毒株间的S基因核苷酸序列同源性为96.9%~100.0%，氨基酸序列同源性为96.1%~100.0%，与22株参考毒株的核苷酸、氨基酸序列同源性分别为92.7%~100.0%和91.5%~100.0%；相较于CV777疫苗株，PEDV江西流行毒株的S基因存在碱基缺失、插入和位点突变现象。本研究从分子流行病学角度证实了2017年江西部分地区PEDV的流行与变异情况，为指导江西省PEDV的科学防控提供了参考依据。

关键词: 猪流行性腹泻病毒(PEDV); 分子流行病学; S基因; 遗传变异

Abstract:

To investigate the epidemiological and genetic variation of porcine epidemic diarrhea virus (PEDV) in Jiangxi province,a total of 182 clinical porcine intestinal and fecal samples were collected from pig farms in some areas of Jiangxi province in 2017 and detected for PEDV by RT-PCR.Two pairs of primers were designed and the S gene from 37 PEDV positive samples were amplified,cloned and sequenced.Genetic evolution analysis of S gene was performed using bioinformatics software and referenced PEDV strains registered in GenBank.The results showed that the length of S gene genome of 37 Jiangxi PEDV strains was 4 158/4 161 bp,encoding 1 385/1 386 amino acids.All PEDV strains belong to Group 1 which had a close relationship with the United States strains,and differed genetically from European strains (Br1/87) and the vaccine strains (CV777).36 of 37 PEDV Jiangxi strains were G1-1 and 1 strain was G1-2.The nucleotides homology of S gene among 37 PEDV strains were 96.9% to 100.0%,and that of amino acids homology was 96.1% to 100.0%.The nucleotides and amino acids homology of S gene between PEDV strains and referenced PEDV strains were 92.7% to 100.0% and 91.5% to 100.0%,respectively.Compared with the vaccine strains (CV777),S gene of PEDV Jiangxi strains existed bases deletions,insertions and mutation.This study confirmed the prevalence and variation of PEDV in Jiangxi province from the perspective of molecular epidemiology,and provided references for guiding the scientific prevention and control of PEDV in Jiangxi province.

Key words: porcine epidemic diarrhea virus (PEDV); molecular epidemiology; S gene; genetic variation

中图分类号:

S852.65

蒋新华, 周荷田, 罗锋, 杨丹凤, 张志庆, 陈松昌, 邓舜洲. 2017年江西部分地区PEDV分子流行病学调查[J]. 《中国畜牧兽医》, 2019, 46(5): 1474-1482.

JIANG Xinhua, ZHOU Hetian, LUO Feng, YANG Danfeng, ZHANG Zhiqing, CHEN Songchang, DENG Shunzhou. Molecular Epidemiological Investigation of Porcine Epidemic Diarrhea Virus in Jiangxi Province in 2017[J]. , 2019, 46(5): 1474-1482.

参考文献

[1] DEBOUCK P,PENSAERT M.Experimental infection of pigs with a new porcine enteric coronavirus,CV777[J].American Journal of Veterinary Research,1980,41(2):219-223.
[2] PENSAERT M B,DE BOUCK P.A new coronavirus-like particle associated with diarrhea in swine[J].Archives of Virology,1978,58(3):243-247.
[3] STEVENSON G W,HOANG H,SCHWARTZ K J,et al.Emergence of porcine epidemic diarrhea virus in the United States:Clinical signs,lesions,and viral genomic sequences[J].Journal of Veterinary Diagnostic Investigation,2013,25(5):649-654.
[4] CHEN J F,SUN D B,WANG C B,et al.Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China[J].Virus Genes,2008,36(2):355-364.
[5] 常铁城,陈建飞,冯力,等.2014年部分地区猪流行性腹泻病毒流行病学调查[J].中国预防兽医学报,2016,38(4):335-338. CHANG T C,CHEN J F,FENG L,et al.Epidemiological survey of porcine epidemic diarrhea virus in 2014[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(4):335-338.(in Chinese)
[6] 李智丽,黄淑坚,马静云,等.2011~2015年我国南方地区PEDV分子流行病学调查及S基因遗传变异分析[J].广东农业科学,2016,43(11):127-135. LI Z L,HUANG S J,MA J Y,et al.Molecular epidemiological investigation of PEDV and analysis of genetic variation of S gene in South China during 2011-2015[J].Guangdong Agricultural Sciences,2016,43(11):127-135.(in Chinese)
[7] 白小磊,崔进,崔甜甜,等.广东部分地区猪流行性腹泻病毒流行病学调查[J].中国兽医学报,2016,36(11):1823-1828. BAI X L,CUI J,CUI T T,et al.Molecular characterization and epidemiological survey of porcine epidemic diarrhea virus in some areas of Guangdong[J].Chinese Journal of Veterinary Science,2016,36(11):1823-1828.(in Chinese)
[8] 赵振鹏,杨振,林伟东,等.江苏省猪流行性腹泻病毒的流行病学调查[J].安徽农业科学,2015,43(19):125-127. ZHAO Z P,YANG Z,LIN W D,et al.Epidemiological investigation of porcine epidemic diarrhea disease in Jiangsu province[J].Journal of Anhui Agricultural Sciences,2015,43(19):125-127.(in Chinese)
[9] 郑逢梅,霍金耀,赵军,等.2010~2012年华中地区猪流行性腹泻病毒分子特征和遗传进化分析[J].病毒学报,2013,29(2):197-205. ZHENG F M,HUO J Y,ZHAO J,et al.Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus field strains in central China during 2010-2012 outbreaks[J].Chinese Journal of Virology,2013,29(2):197-205.(in Chinese)
[10] 董世娟,朱于敏,于瑞嵩,等.新发猪流行性腹泻病毒ORF3基因的遗传进化分析[J].华中农业大学学报,2014,33(5):85-92. DONG S J,ZHU Y M,YU R S,et al.ORF3 gene genetic variation analysis of porcine epidemic diarrhea virus prevailing in China[J].Jorunal of Huazhong Agricultural University,2014,33(5):85-92.(in Chinese)
[11] LEE D K,PARK C K,KIM S H,et al.Heterogeneity in spike protein genes of porcine epidemic diarrhea viruses isolated in Korea[J].Virus Research,2010,149(2):175-182.
[12] HOUDHURY B,DASTJERDI A,DOYLE N,et al.From the field to the lab——An European view on the global spread of PEDV[J].Virus Research,2016,226:40-49.
[13] SONG D,PARK B.Porcine epidemic diarrhoea virus:A comprehensive review of molecular epidemiology,diagnosis,and vaccines[J].Virus Genes,2012,44(2):167-175.
[14] SUN D,FENG L,SHI H,et al.Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein[J].Veterinary Microbiology,2008,131(1-2):73-81.
[15] TIAN P F,JIN Y L,XING G,et al.Evidence of recombinant strains of porcine epidemic diarrhea virus,United States,2013[J].Emerging Infectious Diseases,2014,20(10):1735-1738.
[16] PARK S J,MOON H J,YANG J S,et al.Sequence analysis of the partial spike glycoprotein gene of porcine epidemic diarrhea viruses isolated in Korea[J].Virus Genes,2007,35(2):321-332.
[17] YEO S G,HERNANDEZ M,KRELL P J,et al.Cloning and sequence analysis of the spike gene of porcine epidemic diarrhea virus Chinju 99[J].Virus Genes,2003,26(3):239-246.
[18] 阳酉萍,黄小波,曹三杰,等.四川部分地区株流行性腹泻的分子流行病学调查[J].中国兽医学报,2015,35(5):704-710. YANG Y P,HUANG X B,CAO S J,et al.Molecular epidemiological investigation porcine epidemic diarrhea in some areas of Sichuan[J].Chinese Journal of Veterinary Science,2015,35(5):704-710.(in Chinese)
[19] 张志,程小娜,樊雅婷,等.猪流行性腹泻病毒变异株的鉴定和分子特征分析[J].中国畜牧兽医,2015,42(7):1654-1660. ZHANG Z,CHENG X N,FAN Y T,et al.Identification and molecular characteristics analysis of porcine epidemic diarrhea virus variants[J].China Animal Husbandry & Veterinary Medicine,2015,42(7):1654-1660.(in Chinese)
[20] 谢荣辉,赵灵燕,倪柏峰,等.浙江省猪流行性腹泻病毒S1基因克隆与序列分析[J].中国畜牧兽医,2018,45(6):1668-1676. XIE R H,ZHAO L Y,NI B F,et al.Cloning and sequence analysis of S1 gene of porcine epidemic diarrhea virus from Zhejiang province[J].China Animal Husbandry & Veterinary Medicine,2018,45(6):1668-1676.(in Chinese)
[21] 毛黎红,张双翔,周碧君,等.6株猪流行性腹泻病毒贵州株S基因序列分析[J].中国畜牧兽医,2017,44(6):1621-1629. MAO L H,ZHANG S X,ZHOU B J,et al.Sequence analysis of S gene of 6 strains of porcine epidemic diarrhea virus from Guizhou[J].China Animal Husbandry & Veterinary Medicine,2017,44(6):1621-1629.(in Chinese)
[22] 刘文俊,姚敬明,吴忻,等.猪流行性腹泻病毒山西分离株S基因遗传变异分析[J].中国畜牧兽医,2017,44(7):1982-1989. LIU W J,YAO J M,WU X,et al.Genetic variation analysis of S gene of porcine epidemic diarrhea virus isolated in Shanxi province[J].China Animal Husbandry & Veterinary Medicine,2017,44(7):1982-1989.(in Chinese)