摘要
【目的】确定吉林省某疑似高致病性PRRS猪场的病原,研究其基因特性。【方法】采集疑似感染高致病性PRRSV病猪的脏器组织,处理后接种MARC-145细胞,观察细胞病变,分离PRRSV;应用RT-PCR方法检测和确定分离毒株的基因型,采用间接免疫荧光、电镜观察和全基因组测序检测分离的病毒。【结果】分离毒株为北美洲型PRRSV,命名为JL-04/12,其基因组全长为15 320bp(不包括PolyA),可使MARC-145细胞产生典型的细胞病变。序列比对结果表明:JL-04/12株与经典毒株VR-2332和CH-1a的核苷酸同源性分别为89.5%和94.9%;与高致病性毒株JXA1、HUN4的核苷酸同源性为99.1%。分离毒株JL-04/12编码的2个非结构蛋白和GP2～GP4的氨基酸序列与HUN4株同源性较高,在97.2%～99.3%,而JL-04/12编码的GP5的氨基酸序列与JXA1株同源性较高,为98.5%;JL-04/12编码的GP6和N蛋白的氨基酸序列与高致病性PRRSV毒株JXA1和HUN4的同源性均为100%。PRRSV JL-04/12株的Nsp2不连续缺失30个氨基酸,与高致病性PRRSV毒株JXA1和HUN4的同源性最高,均为97.8%,判定该分离株为变异的高致病性PRRSV。【结论】从吉林省分离到1株高致病性PRRSV,说明高致病性PRRS变异病毒在吉林省仍然存在。
[Objective] The study was to determine the causative pathogen of diseased swine with suspected high pathogenic porcine reproductive and respiratory syndrome in Jilin province, and examine its genetic characteristics. [Method] The tissues infected were disposed and inoculated with MARC-145 ceils. CPE was observed and virus was isolated. Virus detection and genotype identification were conducted by RT-PCR. Then the virus was determined by immunofluorescence, electron microscopy testing and genome sequencing. [Result] The results showed that the isolate was proved to be North American type PRRSV, and named JL-04/12 strain. The full-length of genome was 15 320 bp （PolyA not included）. CPE can be observed on MARC-145 cells. Sequence alignment results indicated that the nucleotide homologies between JL-04/12 strain and classic isolates VR-2332 and CH-la were 89.5% and 94.9% ,respectively while that between both JXA1 and HUN4 were 99.1%. The homologies of the amino acid sequence in two non-structural proteins and GP2--GP4 between JL-04/12 and HUN4 were among 97.2%-99.3%,and that of GP5 between JL-04/12 and JXA1 was 98. 5%. However, GP6 and N protein of JL-04/12 were same as HPPRRSV JXA1 and HUN4 （homology was 100%）. Discontinuous loss of 30 amino acids existed in JL-04/12 Nsp2. Homologies of Nsp2 between both HP-PRRSV JXA1 and HUN4,were 97.8%. Thus,the isolate was a variant of highly pathogenic porcine reproductive and respiratory syndrome. [Conclusion] We obtained an isolate of HP-PRRSV in Jilin, China. It was indicated that high pathogenic porcine reproductive and respiratory syndrome virus existed in Jilin,China.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2013年第4期37-44,共8页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家"863"项目"新型重大动物疫病疫苗与诊断试剂创制及生产工艺创新"(2011AA10A213)
关键词
猪繁殖与呼吸综合征病毒
分离鉴定
全基因组测序
porcine reproductive and respiratory syndrome virus
isolation and identification
genome sequencing
