【作者】 彭昊； 李军； 陶立； 陈泽祥； 唐林生； 韦志锋； 马春霞； 谢宇舟； 禤雄标； 胡帅； 许力干； 谢永平； 杨威； 闭炳芬； 潘艳；

【Author】 PENG Hao1,2,3,LI Jun1,2,TAO Li1,2,CHEN Ze-xiang1,2,TANG Lin-sheng3,WEI Zhi-feng1,2,MA Chun-xia1,2,XIE Yu-zhou1,2,XUAN Xiong-biao1,2,HU Shuai1,2,XU Li-gan1,2,XIE Yong-ping1,2,YANG Wei 1,2,BI Bing-fen1,2,PAN Yan1,2,3(1.Guangxi Veterinary Research Institute,Nanning 530001,China;2 Guangxi Key Laboratory of Animal Vaccines and New Technology,Nanning,53001,China;3.College of Animal Science and Technology,Guangxi University,Nanning 530005,China)

【机构】 广西壮族自治区兽医研究所； 广西畜禽疫苗新技术重点实验室； 广西大学动物科技学院；

【摘要】 根据牛隐孢子虫COWP基因和ITS-1基因,设计合成两对特异性引物,用以特异性扩增牛微小隐孢子虫和安氏隐孢子虫,扩增长度分别为1033bp和263bp。通过反应条件的优化,特异性试验及扩增片段的序列测定的验证,建立了多重PCR方法。应用该方法对采集自广西不同地区的1613份牛粪样进行牛隐孢子虫检测,安氏隐孢子虫阳性粪样为189份,微小隐孢子虫阳性粪样1份,与常规隐孢子虫分离鉴定结果一致。表明该方法可用于隐孢子虫的流行病学调查及临床诊断。更多还原

【Abstract】 Two pairs of specific primer were designed and synthesized according to the specific sequence of COWP and ITS-1 Gene of Dairy Cryptosporidium.A multiplex PCR was developed to amplify specific DNA fragment of C.parvum and C.andersoni with the target sequeuces of 1033bp,263bp.By Optimizing the PCR response condition specific test and the PCR products sequencing,the multi-PCR was established.1613 dairy fecal samples isolated from different areas of Guangxi were tested by the multiplex PCR,then 189 C.andersoni samples and 1 C.parvum sample were detected,which is consistent with conventional Cryptosporidium isolation and identification results.The results showed that the established multi-PCR is suitable for the clinical detection and molecular epidemiology survey of Dairy Cryptosporidium.更多还原