猪流行性腹泻病毒检测与 S 蛋白中和抗原表位基因分析

摘要

应用 RT-PCR 对2015年江苏、安徽、山东9个猪场的腹泻样品（新鲜粪便、肠道组织等）进行 PEDV和 TGV 检测，并对检测到的一株 PEDV（CHNJPK2015）的纤突蛋白中和抗原表位基因进行克隆测序和遗传进化分析。结果表明：9个猪场 PEDV 检测阳性率为100％，PEDV 个体阳性率为63．89％，TGEV 均未检出，说明本次流行的仔猪腹泻病主要是由 PEDV 感染引起的。所测毒株与参考毒株可分为3个群，CHNJPK2015株与AVCT12、Br1？87、CHM2013、LZC、SM98等毒株的 S 蛋白中和抗原表位基因的遗传距离较近，核苷酸序列同源性达99．5％—99．8％，处于同一群；而与2014年中国各地分离的 CH？GDZQ？2014、FJ-FQ 2014、FJ-ZP 2014、PEDV-LY、PEDV-WS、PZ1406、SC1402、SH1404和疫苗株 CV777序列的遗传距离较远，核苷酸序列同源性为92．9％—96.2％，处于不同的群。%Detection of porcine epidemic diarrhea virus (PEDV)and transmissible gastroenteritis virus (TGEV)in diarrhea samples(fresh feces and intestinal tissue,etc.)of 9 pig farms form Jiangsu,Anhui,Shandong in 201 5 were conducted by RT-PCR,and the neutralizing antigen epitope gene of spike protein of isolated PEDV strain(CHNJPK201 5)was cloned,sequenced and genetic evolution analyzed.The results showed that the positive rate of PEDV in 9 pig farms was 1 00%,the individual positive rate of PEDV was 63.89%,no TGEV was detec-ted,indicating that the diarrhea disease was mainly caused by PEDV this time.The tested strain and reference strains could be divided into 3 groups.The neutralizing antigen epitope gene of S protein of CHNJPK201 5 and AVCT1 2,Br1 ?87,CHM201 3,LZC,SM98 strains showed near genetic distance,the nucleotide sequence homology of them were 99.5%—99.8%,belonging to the same group.But the neutralizing antigen epitope gene of S pro-tein of CHNJPK201 5 and CH?GDZQ?201 4,FJ-FQ 201 4,FJ-ZP 201 4,PEDV-LY,PEDV-WS,PZ1 406,SC1 402, SH1 404,vaccine strain CV777 isolated in 201 4 showed far genetic distance,the nucleotide sequence homology of them were 92.9%—96.2%,belonging to different groups.