2株高致病性蓝耳病病毒的分离鉴定及其非结构蛋白Nsp2基因特性分析

Isolation,Identification and Characteristic Analysis of Non-structural Protein Nsp2 Gene of two High Pathogenicity Porcne Reprodutive and Syndrome Virus Strain

关键词:  高致病性蓝耳病病毒,分离,鉴定,Nsp2,特性分析

Key words:  high pathogenicity porcine reproductive and respiratory syndrom e virus  isolation  identification  Nsp2  characteristic analysis

摘要:

为了解黑龙江省猪繁殖与呼吸综合征病毒(PRRSV)的流行现状、分子生物学特征以及遗传演化规律,对黑龙江省不同地区的2个发病猪场采集样品进行了猪繁殖与呼吸综合征病毒的分离鉴定和Nsp2基因遗传进化分析。结果表明,2个猪场的样品均能使Marc-145细胞产生稳定的细胞病变,经RT-PCR鉴定为PRRSV；Nsp2部分基因序列测定发现所扩增的Nsp2基因有30个氨基酸不连续缺失。序列对比结果表明,2个分离株之间的核苷酸同源性98.5%,推导的氨基酸同源性97.4%,与以CH-1a株为代表的疫苗株的核苷酸同源性为77.9%~88.0%,推导的氨基酸同源性为76.7%~86.5% ；与以JXA1株为代表的变异株的核苷酸同源性为96.2%~98.9%,推导的氨基酸同源性为95.4%~97.7%。表明黑龙江省的流行毒株来源于2006~2007年流行毒株,研究结果也为黑龙江省高致病性蓝耳病的流行特点、防控工作提供参考。

Abstract:

To study the current situation, molecular characteristic and genetic evolution of porcine reproductive and respiratory syndrome virus (PRRSV) in Heilongjang provinc，Samples were collected from 2 pig farms in different areas where suffering the disease for virus isolation, identification and analysis of the Nsp2 Gene. The results showed that two PRRSV stain were isolated and identified through the RT-PCR assay, and the 30 discontinuous deletion amio acids were found in the Nsp2 gene. Sequence analysis idicated that homology between the two strain were 98.5% in nucleotide and 97.4% in amino acids sequence; the homology between the two strain and the classical strain, CH-1a strain for example, were 77.9%-88.0% in nucleotide and 76.7%-86.5% in amino acids sequence; the homology between the two strain and the HP-PRRS variation, JXA1 strain for example, were 96.2%-98.9% in nucleotide and 95.4%-97.7% in amino acids sequence. It was indicated that the epidemical strain in Heilongjiang province originated from popular strain 2006-2007, and a reference for the epidemic characteristic and the disease preventon and control work for the HP-PRRSV in Heiongjiang Province were provided, also.