中国兽医杂志2025，Vol.61Issue(8)：46-52,7.DOI:10.20157/j.cnki.zgsyzz.2025.08.007
牛白血病病毒RAA荧光检测方法的建立与初步应用
Establishment and Preliminary Application of RAA Fluorescent Detection Method for Bovine Leukemia Virus
摘要
Abstract
To establish an efficient,convenient,and specific recombinase-mediated isothermal amplification(RAA)rapid detection method based on the env structural gene of bovine leukemia virus(BLV),this study used DNAMAN for sequence alignment,constructed the recombinant plasmid pESI-env,designed and screened RAA primers and probes,established the reaction system,and further optimized the reaction protocol and primer-probe usage.The method's specificity,sensitivity,and reproducibility were evaluated,and the results were compared with enzyme-linked immunosorbent assay(ELISA)and quantitative PCR(qPCR)in clinical samples.The results showed that the optimal primer-probe combination was F4/R1/P.The method completed the reaction in 20 minutes.There was no cross-reaction with bovine viral diarrhea virus(BVDV),infectious bovine rhinotracheitis virus(IBRV),and bovine rotavirus(BRV).The minimum detection limit was 1.62×101 copies/μL,and the method showed good repeatability.The positive detection rate of clinical samples using this method was slightly higher than that of both ELISA and qPCR standard methods.This study successfully established an efficient,convenient,and highly specific BLV RAA fluorescent detection method,which eliminates the reliance on expensive equipment,specialized personnel,and complex detection techniques,providing technical support for BLV clinical detection.关键词
牛白血病病毒(BLV)/重组酶介导等温扩增(RAA)/快速检测方法Key words
bovine leukemia virus(BLV)/recombinase aided amplification(RAA)/rapid detection method分类
农业科技引用本文复制引用
侯欣怡,王建发,连帅,耿子健,武瑞..牛白血病病毒RAA荧光检测方法的建立与初步应用[J].中国兽医杂志,2025,61(8):46-52,7.基金项目
国家自然科学基金项目(31972747) （31972747）
