鹅细小病毒TaqMan荧光定量PCR方法的建立及临床应用  

Establishment and clinical application of TaqMan fluorescent quantitative PCR assay for detection of goose parvovirus

作　　者：王艳秋 张紫璇 高旭[1] 李卓昕 王美琪 刘晋宇[1] 鲁承[1] 梁晚枫[1] 于龙政[1] 王妍[1] WANG Yan-qiu;ZHANG Zi-xuan;GAO Xu;LI Zhuo-xin;WANG Mei-qi;LIU Jin-yu;LU Cheng;LIANG Wan-feng;YU Long-zheng;WANG Yan(Agricultural College ,Yanbian University ,Yanji, Jilin 133400, China)

机构地区：[1]延边大学农学院

出　　处：《中国兽医学报》2018年第6期1100-1104,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31460661)

摘　　要：为了建立一种能定量检测鹅细小病毒（GPV）的荧光定量PCR方法，根据GenBank已收录GPVVP3基因保守区设计1对特异引物和TaqMan探针，经反应条件优化、标准曲线建立，以及敏感性、特异性和重复性试验。结果显示，该方法在10^7～10^2拷贝/μL具有良好的线性关系（R^2=0．999）；灵敏度是普通PCR方法的100倍；对其他3种常见禽病毒无特异性扩增，具有较好的特异性；组内与组间的变异系数均小于2％，具有良好的重复性。对临床20份疑似GPV感染鹅病料进行检测，检出率比普通PCR高10％。表明本试验建立的TaqMan荧光定量PCR方法准确、稳定、灵敏和特异，可以用于GPV临床定性与定量检测。In order to establish a TaqMan quantitative fluorescent PCR method for quantitative de tection of goose parvovirus. A pair of specific primers and probes were designed according to the conserved region of GPV-VP3 gene collected in GenBank, the reaction conditions were optimized, the standard curve was established, and the sensitivity, specificity and repeatability of the method were validated. The results showed that the proposed method has good linear relationship at 10^7-10^2 copies /μL, sensitivity is 100 higher times than the normal PCR method, there is no specific amplification of the other three common avian viruses, the coefficient of variation between group and group was less than 2 %. The clinical data of 20 suspected GPV infected goose were tested and the detection rate was 10%, higher than that of common PCR, These results indicated that the TaqMan fluorescent quantitative PCR method established in this study was accurate, sensitive and specific,and can be used for clinical qualitative and quantitative detection of GPV.

关 键 词：鹅细小病毒 VP3基因 实时定量PCR TaqMan荧光探针 

分 类 号：S852.65[农业科学—基础兽医学]