禽多瘤病毒TaqMan探针实时荧光定量PCR检测方法的建立及应用  

Establishment and application of TaqMan probe real-time fluorescent quantitative PCR for detection of avian polyomavirus

作　　者：闫佳佳 翟天舒 许冠龙 王嘉[2] 孔冬妮[2] 邓永[2] 汪溪 薛青红[2] 程佳[1] 毛娅卿[2] 印春生[2] YAN Jia-jia;ZHAI Tian-shu;XU Guan-long;WANG Jia;KONG Dong-ni;DENG Yong;WANG Xi;XUE Qing-hong;CHENG Jia;MAO Ya-qing;YIN Chun-sheng(College of Veterinary Medicine,Shanxi Agricultural University,Taigu 030801,China;China Institute of Veterinary Drug Control,Beijing 100081,China;College of Animal Science and Technology,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]山西农业大学动物医学学院,山西太谷030801 [2]中国兽医药品监察所,北京100081 [3]江西农业大学动物科学技术学院,江西南昌330045

出　　处：《中国兽医科学》2023年第5期566-571,共6页Chinese Veterinary Science

基　　金：国家自然科学基金青年科学基金项目(32002273)。

摘　　要：为建立检测禽多瘤病毒(avian polyomavirus,APV)的快速诊断方法,根据APV VP4基因的保守序列设计并合成引物和探针,建立了TaqMan探针实时荧光定量PCR检测方法,验证了其特异性、敏感性和重复性,并利用建立的方法对采集的临床样本组织进行检测。结果显示,建立的TaqMan探针实时荧光定量PCR检测方法的C_(t)值与标准品在1.0×10^(9)~1.0×10^(2)copies/μL范围内呈良好的线性关系,相关系数为0.996,斜率为-3.021,检测下限为1.0×10^(1)copies/μL,与其他禽源病毒未发生交叉反应,重复性试验的变异系数均小于2%,重复性好,并且该方法在实际临床检测中可行。上述结果表明,建立的方法可用于禽多瘤病的早期诊断和APV快速检测。Development of a rapid diagnostic method for detecting avian polyomavirus(APV).Primers and probes were designed and synthesized according to the conserved sequence of VP4 gene of APV.A TaqMan probe real-time fluorescence quantitative PCR detection method was established,and its specificity,sensitivity and repeatability were verified.The collected clinical sample tissues were detected by the established method.Result,the C_(t) value of the established Taq Man probe real-time fluorescent quantitative PCR detection method showed a good linear relationship with the standard in the range of1.0×10^(9)—1.0×10^(2) copies/μL.The correlation coefficient was 0.996,the slope was-3.021,and the detection limit was 1.0×10^(1)copies/μL.There was no cross-reaction with other avian viruses.The coefficient of variation of the repeatability test was less than 2%,and the repeatability was good.The method was feasible in actual clinical detection.This method can be used for early diagnosis and rapid detection of APV.

关 键 词：禽多瘤病毒 VP4基因 TaqMan探针实时荧光定量PCR 

分 类 号：S852.659.1[农业科学—基础兽医学]