| 猪圆环病毒2型毒株分离鉴定和体外增殖特性研究 |
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| 引用本文： | 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. |
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| 作者姓名： | 王贵华 陈义锋 汤波 刘明明 陈翠云 赵亚荣 |
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| 作者单位： | 北京大北农动物医学研究中心,北京,100097 |
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| 摘 要： | 为分离鉴定猪圆环病毒2型(PCV2)毒株并研究其体外增殖特性,本研究采用PCR法分别对从山东、山西、北京和河北收集的疑似断奶仔猪多系统衰竭综合征(PMWS)病猪的病料进行检测,将检测结果为PCV2阳性的病料处理后接种PCV阴性的PK15A细胞连续传代进行病毒分离,观察每代细胞是否有病变,采用PCR、间接免疫荧光法(IFA)对第3代培养物进行鉴定,对各毒株的全基因组进行克隆、测序和系统进化分析,并将PCV2各分离株连续传20代,对其体外增殖特性进行了研究。结果共分离到5株猪圆环病毒2型毒株,分别命名为DBN-SD02、DBN-SX07 DBN-BJ08、DBN-HB12和DBN-HB13株,GenBank登录号分别为FJ660967、FJ660968、FJ660969、FJ660970和FJ660971。各分离株基因组全长均为1767 bp,系统进化分析结果表明,除DBN BJ-08属于PCV2a基因型外,其余均属于PCV2b基因型,基因组序列核苷酸同源性为98.5%~99.7%,体外增殖特性研究结果表明,随着传代次数的增加病毒感染滴度均有明显的提高。
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| 关 键 词： | 猪圆环病毒2型 分离 鉴定 增殖特性 |
Research on Isolation, Identification and Characteristic of Vitro Propagation of Isolates of Porcine Circovirus Type 2 |
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| WANG Gui-hua
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CHEN Yi-feng
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TANG Bo
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LIU Ming-ming
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CHEN Cui-yun
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ZHAO Ya-rong.Research on Isolation, Identification and Characteristic of Vitro Propagation of Isolates of Porcine Circovirus Type 2[J].China Animal Husbandry & Veterinary Medicine,2012,39(1):17-22. |
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| Authors: | WANG Gui-hua CHEN Yi-feng TANG Bo LIU Ming-ming CHEN Cui-yun ZHAO Ya-rong |
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| Institution: | Veterinary Medicine Research Center of Beijing Da Bei Nong Group,Beijing 100097,China |
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| 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(TCID50) 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 TCID50 increase obviously with the times of propagation. |
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| Keywords: | porcine circovirus type 2 isolation identification characteristic of propagation |
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