动物医学进展2025，Vol.46Issue(8)：7-11,5.
山羊星状病毒TaqMan荧光定量PCR检测方法的建立与应用
Establishment and Application of TaqMan Fluorescence Quantitative PCR Detection Method for Caprine Astrovirus
摘要
Abstract
In order to establish a rapid detection method for caprine astrovirus,specific probe and primers were designed referring to the ORF1ab gene sequence of caprine astrovirus(OR652457)collected in Gen-Bank,and the recombinant plasmid was constructed.The recombinant plasmid was employed as the stand-ard substance to optimizing the reaction conditions and the preliminary application of clinical sample detec-tion,the TaqMan fluorescence quantitative PCR method for detection of caprine astrovirus was established.The results indicated that the standard curve established by this method has good linear relationship.There was no cross reaction against BVDV,BCoV,BRV,Coxiella burnetii and Mycoplasma.The minimum detec-tion limit of recombinant plasmid of caprine astrovirus standard substance was 2.16×101 copies/μL.The results of intra-batch and inter-batch tests were stable,and the coefficient of variation was less than 2％.The method established in this study was used to detect 115 diarrhea stool samples collected from some goat farms in Shaanxi province.The results indicated that the positive rate of caprine astrovirus was 29.57％,and the coincidence rate with the conventional RT-PCR detection was 94.78％.This method pro-vides a reference technique for the rapid detection of caprine astrovirus.关键词
山羊星状病毒/TaqMan荧光定量PCR/检测方法Key words
Caprine astrovirus/TaqMan real-time PCR/Detection method分类
农业科技引用本文复制引用
于皓同,马勇杰,王凯茸,张琪,张淑霞,许信刚..山羊星状病毒TaqMan荧光定量PCR检测方法的建立与应用[J].动物医学进展,2025,46(8):7-11,5.基金项目
国家重点研发计划项目(2021YFD1600704) （2021YFD1600704）
陕西省农业专项资金项目(XNDY2210) （XNDY2210）
