犬细小病毒PCR诊断方法的建立及对大熊猫粪便的检测  

Establishment of PCR for detection of canine parvovirus and investigation into gi ant pandas

作　　者：郭玲[1] 杨绍林[1] 张志和 陈世界[3] 阳爱国[4] 李德生[5] 王成东[5] 刘丹[1] 刘杰[1] 潘海波[1] 李乡城[1] 舒蕾[1] 颜其贵[1,6] 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]成都大熊猫繁育研究基地,四川成都610081 [3]四川出入境检验检疫局,四川成都610041 [4]四川省动物疫病预防控制中心,四川成都610041 [5]中国保护大熊猫研究中心,四川雅安625000 [6]四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医学报》2013年第8期1201-1205,共5页Chinese Journal of Veterinary Science

基　　金：科技部公益性行业科研专项资助项目(200910188);四川省科技厅科技支撑计划资助项目(2010SZ0050);国家林业公益性行业科研专项资助项目(201104050);国家"973"计划前期研究专项资助项目(2012CB722207)

摘　　要：根据GenBank中登录的犬细小病毒(CPV)VP2基因序列,设计合成1对特异性引物;以CPV疫苗株为模板,建立了一种快速检测CPV的PCR检测方法,并应用于CPV诊断。结果显示,以此对引物进行PCR扩增能得到与理论设计值大小一致的342bp的特异性条带,对犬瘟热病毒(CDV)、犬冠状病毒(CCV)、狂犬病病毒(RV)、新城疫病毒(NDV)和猫泛白细胞减少症病毒(FPV)扩增结果均为阴性;最低可检出约1.4pg的病毒核酸;重复性试验结果表明,其检测重复性好;对45份临床宠物犬病料进行检测,并与免疫胶体金抗原检测拭纸检测结果进行比较,吻合率为90.0%。将此方法初步应用于大熊猫粪便中细小病毒的检测,结果表明,从熊猫基地采集的52份正常大熊猫粪便样品中有8份为细小病毒阳性,阳性率为15.3%。大熊猫是通过自然感染还是弱毒苗感染细小病毒的机制还不清楚。Based on the published sequence of the VP2 gene of canine parvovirus（CPV） in Gen- Bank,a pair of specific primers was designed for the detection of CPV. As a template of the CPV vaccine strain,and after optimization of the reaction system, a rapid PCR detection method for CPV was established. A expected fragment of 342 bp was amplified from the template. As little as 1.4 pg DNA of CPV could be detected accurately and rapidly. This method had good reproducibili-ty and stability. There was no cross reaction to canine distemper virus, canine coronavirus, rabies virus, Newcastle disease virus and feline panleukopenia virus. Suspected disease materials had been tested by PCR, and the PCR results were compared with immune colloidal gold technique and the coinci Research Center for the dence rate was up to 90.0%. Eight of the 52 normal giant pandas from the Giant Panda were detected infection with the virus, the morbidity rate was 64.7 %. The route of infection with these pathogens via natural exposure or from m cines in giant pandas is unknown. The results of this study emphasize the need for fectious diseases of giant pandas. odified live vac-research on in-

关 键 词：犬细小病毒 PCR 大熊猫 

分 类 号：S852.65[农业科学—基础兽医学]