摘要
用RT-PCR方法从某猪场病猪体内分离到1株猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV),命名为PRRSV GD-ZQ株,可致Marc-145细胞病变。细胞分离培育病毒至增殖性能稳定后,分别扩增分离病毒的Nsp2、ORF3、ORF5基因并测序,绘制进化树比较分析遗传进化。结果显示分离毒株的Nsp2、ORF3、ORF5基因序列与美洲型VR-2332、CH-1a等毒株核苷酸序列相似性在89.0%～95.8%之间,与欧洲毒株LV的相似性在48.2%～49.3%之间,遗传进化分析显示该毒株与CH-1a株处于同一分支,分离毒株的Nsp2序列与以HEB1为代表的PRRSV变异株相似性在98.3%～99.4%之间。该病毒在细胞盲传5代能产生稳定规律细胞病变(CPE),且第5代病毒TCID50达10-5.6/0.1 mL。进化分析结果显示GD-ZQ分离株是PRRSV变异株,具有"高热病"PRRSV毒株的变异特点,为了解PRRSV在时间上和分子水平上的遗传变异规律、变异幅度和变异范围奠定基础。
Using RT-PCR identification,we obtained a PRRSV variant strain GD-ZQ,which came from the suspected porcine reproductive and respiratory syndrome areas.The variant strain could cause significant CPE in Marc-145 cell.Serial passage of GD-ZQ until it was stable,Nsp2,ORF3,ORF5 gene of the strain were amplified,the sequence determination,the comparative analysis and the cladogram was drawn up.The results indicated the amino acid sequence homology of the three GD-ZQ genes were 89.0% to 95.8% with the American VR-2332 and Ch-1a,and 48.2% to 49.3% with European LV.Further comparative analysis showed GD-ZQ was in the same branch with Ch-1a,amino acid sequence homology of GD-ZQ isolates Nsp2 was 98.3% to 99.4% with other variant strains as HEB1.The 5th generation was could cause regular CPE,with TCID50 of 10-5.6/0.1 mL.The analysis explained that GD-ZQ was a variant strain,with the characteristic of ＂high fever＂ disease mutation of PRRSV strain.The results layed a foundation for the further research on PRRSV in time and molecular level to understand genetic variation and the range of variation.
出处
《中国畜牧兽医》
CAS
北大核心
2011年第5期121-125,共5页
China Animal Husbandry & Veterinary Medicine
