云南省猪圆环病毒2型遗传进化分析

    Genetic Evolution Analysis of Porcine Circovirus Type 2 in Yunnan Province

    • 摘要:
      目的  调查猪圆环病毒2型(PCV2)在云南猪群和牛群中的流行情况,明确云南PCV2毒株的遗传变异、分子进化趋势。
      方法  采用PCR方法对云南省11个地区783份猪、牛的血液、粪便等样品进行PCV2病原学检测,对6份PCV2阳性样品进行全基因组扩增、克隆与序列测定,利用DNAStar软件比对分析全基因和ORF2氨基酸序列,使用MEGA软件绘制遗传进化树。
      结果  检测样品的PCV2阳性率为48.73%;共测得6株PCV2全基因组序列,其中5株为1 767 bp, 1株为1 768 bp;经遗传发育分析可知,有PCV2a亚型2株、PCV2b亚型2株、PCV2d亚型2株;6株PCV2毒株同源性在94.6%~99.9% 之间,与GenBank数据库中20条参考株同源性在91.4%~99.8% 之间;6株PCV2亚型的ORF2都有其典型的氨基酸位点,PCV2a毒株在第80(V80L) 氨基酸位点突变为与PCV2(b/d)一致,PCV2b毒株在59(R59K)、63(K63R)、190(A190T)等氨基酸位点突变与PCV2d一致。
      结论  PCV2在云南猪群中普遍存在,以PCV2a、PCV2b和PCV2d这3个基因亚型共存为主,且PCV2(a/b)有向PCV2d突变的趋势,荷斯坦奶牛尚未感染PCV2。

       

      Abstract:
      Objective  The study was carried out to investigate the prevalence of porcine circovirus Type 2 (PCV2) in pigs and cattle in Yunnan Province, and to identify the genetic variation and molecular evolution trend of PCV2 strains in Yunnan Province.
      Method  PCR method was used to detect the etiology of PCV2 in 783 blood and manure samples of pigs and cattle from 11 regions of Yunnan Province. The whole genome amplification, cloning and sequencing of 6 PCV2 positive samples were carried out. The whole gene and ORF2 amino acid sequences were compared and analyzed by DNAStar software, and the genetic evolution tree was drawn by MEGA software.
      Result  The positive rate of PCV2 of detected samples was 48.73%. A total of 6 strain PCV2 genome sequences were measured, of which 5 strain were 1 767 bp and 1 strain was 1 768 bp. The genetic development analysis showed that there were 2 strains of PCV2a subtype, 2 strains of PCV2b subtype and 2 strains of PCV2d subtype. The homology of 6 strains PCV2 was 94.6%-99.9%, and the homology with 20 national reference strains in GenBank database was 91.4 %-99.8 %. The ORF2 of 6 strains PCV2 had their typical amino acid sites. The amino acid site of PCV2a strain was mutated to be consistent with PCV2 (b/d) at the 80th (V80L) amino acid site, and the amino acid site of PCV2b strain was mutated to be consistent with PCV2d at the 59th (R59K), 63rd (K63R), 190th (A190T) and other amino acid sites.
      Conclusion  PCV2 commonly exists in pigs in Yunnan, and the three gene subtypes of PCV2a, PCV2b and PCV2d coexist, and PCV2 (a/b) has a tendency to mutate to PCV2d. Holstein cows have not been infected with PCV2.

       

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