畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (6): 717-725.doi:

• 预防兽医 • 上一篇    下一篇

猪繁殖与呼吸综合征病毒TP株P4、P60、P90基因组遗传变异分析及动物致病力试验

招丽婵,苏润环,邓雨修,王东东,杨明柳,李春梅,罗小飞,宋延华*   

  1. 广东温氏食品集团有限公司研究院,新兴 527400
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-06-25 发布日期:2010-06-25
  • 通讯作者: 宋延华

Genetic Variation Analysis and Animal Pathogenicity Test for PRRSV TP Strain P4, P60, P90 Genome

ZHAO Li-chan, SU Run-huan, DENG Yu-xiu, WANG Dong-dong,YANG Ming-liu, LI Chun-mei, LUO Xiao-fei, SONG Yan-hua*   

  1. Guangdong Wen′s Foodstuffs Group Co.LTD, Guangdong Wen′sGroup Academy, Xinxing 527400, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-06-25 Published:2010-06-25

摘要: 本研究旨在对猪繁殖与呼吸综合征病毒(PRRSV) TP 株P4、P60、P90三个代次毒株进行全基因组序列测定,分析其遗传变异特性。通过分段RT-PCR、5′RACE和3′RACE法扩增病毒基因组,并运用生物信息学软件和动物致病力试验进行相关分析。结果表明:TP株P4、P60、P90株基因组全长分别为15 346、15 394和15 393 bp。3个代次病毒的基因组序列中ORF1a基因和3′UTR区域变异较大,尤以Poly(A)尾序列碱基数差异显著,分别为26、74和73 bp。3代次毒株基因组序列比对发现TP P90与TP P4差异较大,共有58个核苷酸位点和27个氨基酸位点发生突变,P90 GP5蛋白抗原指数和表面亲和力与TP P4和TP P60相比呈增加趋势,动物致病力试验表明TP P4毒株致病力较TP P60和 TP P90强。结果提示:TP P4、P60、P90 3代次毒株基因组序列存在着一定的差异,主要集中在4~60代致弱阶段,遗传进化关系上该病毒分属于PRRSV美洲株分支,并与高致病性PRRSV JXA1株亲缘关系最为接近,相似性高达99.5%、99.2%和99.3%。

Abstract: The aim of this experiment was to study the genetic variability of the three different generations of porcine reproductive and respiratory syndrome virus (PRRSV) TP strain, P4, P60, P90. RT-PCR, 5′ RACE and 3′ RACE were employed to amplify the virus genome of the three generations of PRRSV TP strain, and the genomic analyses were conducted using some bioinformatics software including the clustalx and DNAStar, and the animal pathogenicity test. The sequences of the TP strain P4, P60, P90 were 15 346, 15 394 and 15 393 bp, respectively. The variations of the ORF1a gene and 3′UTR were obvious in these three generations genome sequences, especially the poly(A) tails in the 3′UTR region,they were 26, 74 and 73 bp, respectively. Comparing the genome sequences of TP P90 strain with TP P4 strain, there found 58 nucleotide mutations that resulted in 27 amino acid changes. The antigenic index and surface probability of GP5 protein of TP P90 strain were increasing than P4 and P60. Moreover, the virulence test indicated that the pathogenicity of TP P4 is stronger than the TP P60, P90 strain. The results showed that the genome sequences of the three different generations has some variations, especially between passage 4 and 60. The PRRSV TP P4, P60, P90 all belong to the PRRSV America strain branch, and display the highest nucleotide identity to the high pathogenic PRRSV JXA1 strain, 99.5%, 99.2% and 99.3%, respectively.