犬细小病毒巢式PCR诊断方法的建立及应用  

Development and Application of Nested-PCR Method for Detection of Canine Parvovirus(CPV)from Pet Dogs in Xinjiang

作　　者：简子健[1] 马素贞[1] 陈胜男[1] 翟少华[1] 赵森[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业科学》2012年第1期155-160,共6页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区高技术项目(200611107)

摘　　要：【目的】以期建立犬细小病毒巢式PCR诊断方法。【方法】根据基因库已发表的CPV VP2基因序列设计二对巢式引物,以临床上疑似CPV感染的犬粪样品中提取的总DNA作为模板,用巢式PCR方法扩增出CPV VP2基因的目的条带。【结果】扩增出的目的基因与国际标准序列的同源性为99.3%～100%。特异性试验发现该引物不能启动犬其它常见病毒基因的扩增。敏感性试验表明该引物能检测到10-9的CPV总RNA量。重复性试验显示该引物具有良好的稳定性。对采集的26份样品进行CPV巢式PCR检测,20份样品为阳性,阳性率为76.9%。【结论】建立的CPV的RT-PCR检测方法可于临床CPV的快速诊断和小规模的分子流行病学调查。[ Objective and Method ] In order to establish the nested PCR for detection of CPV, two pairs of the special primers were designed according to the VP2 gene sequence of CPV published in GenBank and the specific target stripe was found in the total DNA template from the stool samples of clinically infected dogs with CPV. [ Result] Sequence analysis was indicated which shared 99.3% -100% nueleotide homology with that published in GenBank. Specific experiment of nested PCR was showed the primers could only amplify the genome of CPV, rather than those of other canine viruses. As little as 10 - 9 total DNA template of CPV could be detected in the sensitive test. Reproducible test was highly stability. 20 were positive for CPV infection from 26 tested samples and the prevalence of CPV from the tested dogs was 76. 92% by this nested PCR. [ Conclusion] The established nested- PCR method for detection of the canine parvovirus in this study could be used in the early clinical diagnosis of CPV and the small - scale of epidemiological survey in molecular level.

关 键 词：犬细小病毒 巢式PCR 检测 应用 

分 类 号：S852.655[农业科学—基础兽医学]