›› 2013, Vol. ›› Issue (7): 38-42.

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Isolation,Identification and Genome Sequence Analysis of Porcine Circovirus Type Ⅱ Isolate

YANG Guo-liang, SUN Feng-ting, ZHENG Jie, LONG Jin-xue, ZHANG Tan, ZHANG Hong, DENG Qiu-hong, WANG Ya-li, ZHANG Lin-yun, WANG Wen-quan   

  1. Daxing Biological and Pharmaceutical Industrial Base, Beijing Ceva Huadu Biological Products Co., Ltd., Beijing 102600, China
  • Received:2012-12-13 Online:2013-07-20 Published:2013-07-24

Abstract: In November 2011, an infectious disease characterized by systemic macular hemorrhage, diarrhea and obviously inguinal lymph node enlargement, broke out in a swine farm located in Beijing. The lymph node, kidney and spleen tissues were taken from infected pigs and grinded to obtain tissue suspension which was then filtered through 0.2 μm filter and inoculated into the PK15 cells. D-glucosamine was added to the infected cell which was then gone through 3 blind passages,and DNA was extracted from the harvested virus suspension. Identification were done by PCR method and immunofluorescent assay,and the complete genome was also amplified and sequenced,which was then analyzed by using DNAStar software. The results demonstrated that the isolated strain belonged to PCV2 (named BJ20111113 strain) with whole genome size of 1767 bp. At nucleotide sequence level, its homology could reach 94.5% to 99.8% compared with the domestic and foreign reference strains. Among them, the most closely related strain was HM038017 (BDH strain) with homology of 99.8%, and the far distantly related strain was EU148504 with homology of only 94.5%. The phylogenetic analysis result indicated that BJ20111113 strain belonged to genotype PCV2b.

Key words: porcine circovirus typeⅡ; isolation and identification; sequence analysis

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