中国畜牧兽医2012，Vol.39Issue(1)：17-22,6.
猪圆环病毒2型毒株分离鉴定和体外增殖特性研究
Research on Isolation, Identification and Characteristic of Vitro Propagation of Isolates of Porcine Circovirus Type 2
王贵华 1陈义锋 1汤波 1刘明明 1陈翠云 1赵亚荣1
作者信息
- 1. 北京大北农动物医学研究中心,北京100097
- 折叠
摘要
Abstract
To isolate and identify porcine circovirus type 2 (PCV2)strains,and investigate its' characteristic of vitro propagation, polymerase chain reaction (PCR) was utilized in this study to detect the specimens of suspected PMWS cases collected from Shangdong, Shangxi, Beijing and Hebei province, the positive samples were dealt with and inoculated into the PCV-free PK15A cells which were then propagate continuously to isolate PCV2 strains. The cytopathic effect (CPE) was examined each time and the cultures of the 3rd time were identified by PCR and indirect immunofluorescence assay (IFA). The genome of each isolates were then cloned and sequenced. The sequence was submitted to the GenBank. Based on the sequence phylogenetic tree analysis was made too, subsequently these PCV2 strains' characteristic of vitro propagation were studied by propagated continuously to the 20th generation, and the 5th, 10th, 15th and 20th generation's titer known as 50% tissue culture infectious dose (TCID;o ) were detected These results showed that there were five strains of PCV2 were isolated and identified named PCV2 DBN-SD02,DBN-SX07,DBN-BJ08, DBN-HB12 and DBN-HB13 respectively, and the GenBank submission number were FJ660967, FJ660968, FJ660969, FJ660970.FJ660971,respectively. The genome of five isolates was all 1767 bp. Four of five isolates were belong to the PCV2b genotype and share a 98. 5 % - 99. 7% identity except PCV2 DBN-BJ08 strain which belong to the PCV2a genotype. The vitro propagation study showed that the TCIDfeo increase obviously with the times of propagation.关键词
猪圆环病毒2型/分离/鉴定/增殖特性Key words
porcine circovirus type 2/ isolation/ identification/ characteristic of propagation分类
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王贵华,陈义锋,汤波,刘明明,陈翠云,赵亚荣..猪圆环病毒2型毒株分离鉴定和体外增殖特性研究[J].中国畜牧兽医,2012,39(1):17-22,6.
