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.
作者
闫佳佳
翟天舒
许冠龙
王嘉
孔冬妮
邓永
汪溪
薛青红
程佳
毛娅卿
印春生
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)
出处
《中国兽医科学》
CAS
CSCD
北大核心
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)。
作者简介
闫佳佳(1997-),女,山西山阴人,硕士生,E-mail:yanjiajia1997@163.com;通讯作者:程佳(1978-),女,山西清徐人,副教授,博士,主要从事兽医药理学与毒理学的教学与科研工作,E-mail:sxtgchengjia@163.com;通讯作者:毛娅卿(1981-),女,江西上饶人,研究员,博士,研究方向为禽用病毒性兽用生物制品及检验,E-mail:wcpmyq@163.com;通讯作者:印春生(1977-),男,江苏响水人,研究员,博士,研究方向为兽医微生物与免疫、兽用生物制品检验及相关标准物质的研制,E-mail:zjs62158844@sohu.com。
