| 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15 320 bp（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-4/12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。 |
| 关键词: 猪繁殖与呼吸综合征病毒 分离鉴定 全基因组测序 |
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| 基金项目: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) |
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| Isolation,identification and sequencing of PRRSV JL-04/12 strain with high pathogenicity from Jilin,China |
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WANG Fengxue, GUO Li, WEN Yongjun,et al
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Institute of Special Economic Animal and Plant Science,Chinese Academy of Agricultural Sciences
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| Abstract: |
| 【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 cells.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-1a 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 HP-PRRSV 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. |
| Key words: porcine reproductive and respiratory syndrome virus isolation and identification genome sequencing |
