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2013-2014年我国东北地区猪繁殖与呼吸综合征病毒GP5和NSP2基因遗传变异分析
常晓博,张辉
0
(中国农业科学院哈尔滨兽医研究所;吉林农业大学 动物科学技术学院)
摘要:
为了解2013-2014年我国东北地区猪繁殖与呼吸综合征病毒(PRRSV)的遗传变异情况,研究对东北地区3个省份29个地区采集的123份样品进行了检测,并对20份阳性样品的GP5和部分NSP2基因进行了RT-PCR扩增、测序、遗传进化及序列比对分析。结果表明:我国东北地区PRRSV阳性率为16%,与国内外已发表的10株PRRSV参考株相比,GP5基因的核苷酸同源性为84.4%~100%。GP5氨基酸序列分析表明,大多数毒株在氨基酸高变区、中和表位和诱导表位发生了变异,主要变异为:C24→Y24,F25→L25,V27→A27,L28→I28,N34→G34/S34/D34。LN283株GP5和NSP2基因遗传进化树的不同结果表明,LN283株可能涉及PRRSV基因重组。HLJ13株NSP2基因PCR扩增产物有2条相差90 bp的目的条带,且NSP2基因遗传进化树显示两条序列分别属于CH-1a亚群和HuN4亚群,推断HLJ13株可能为经典和高致病性混合感染株。GP5和NSP2遗传进化及氨基酸序列分析结果表明,所有病毒株均属于北美洲型,且以HP-PRRSV为主,经典株与变异株共存。本研究结果可为我国东北地区PRRSV遗传进化研究及疫病防控提供参考。
关键词:  PRRSV  GP5  NSP2  遗传变异
DOI:
投稿时间:2015-04-14修订日期:2015-07-09
基金项目:国家自然科学基金(31270045);吉林省科技厅自然科学基金项目(20150101107JC)
Genetic Variation Analysis of GP5 and NSP2 of PRRSV in Northeast China from 2013 to 2014
(State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin)
Abstract:
To investigate the genetic variation of pathogenic porcine reproductive and respiratory syndrome(PRRSV) in Northeast China included 29 regions of 3 provinces from 2013 to 2014,123 tissue samples collected from Northeast China were tested, the GP5 and partial NSP2 genes of 20 positive samples were amplified, sequenced, phylogenetic and alignment analyzed, respectively. The results showed that the PRRSV positive rate was 16%, GP5 genesshared 84.4%~100% nucleotide similarity compared with 10 domestic and overseas reference PRRSV strains. Amino acid analysis of GP5 revealed that most strains had mutations, especially in the hyper-variable regions, the primary neutralizing epitope and decoy epitope, which were C24→Y24,F25→L25,V27→A27,L28→I28,N34→G34/S34/D34. The phylogenetic of GP5 and NSP2 of LN283 indicated the strain could be genetic recombination strain.The result of PCR on HLJ13 showed NSP2 had two genes which differed 90 bp, and the phylogenetic of NSP2 showed they belonged to CH-1a and HuN4 subgroup, which could be polyinfection involved of classical and highly pathogenic strain. The phylogenetic and amino acid analysis of GP5 and NSP2 demonstrate all strains belonged to North American genotype, which gave priority to HP-PRRSV, coexisting of classic strains and mutant strains. The study would provide a new reference on genetic evolution and disease control for PRRSV in Northeast China.
Key words:  PRRSV  GP5  NSP2  genetic variation

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