中国动物检疫2025，Vol.42Issue(6)：92-98,7.DOI:10.3969/j.issn.1005-944X.2025.06.015
猪圆环病毒2型和弓形虫双重TaqMan荧光定量PCR法的建立及应用
Establishment and Application of a Dual TaqMan Fluorescence Quantitative PCR Assay for Simultaneous Detection of Porcine Circovirus Type 2 and Toxoplasma gondii
摘要
Abstract
It has been difficult to accurately identify porcine circovirus type 2(PCV2)infection and toxoplasmosis due to their similarity in clinical symptoms.To this end,specific primers and TaqMan probes were designed based on PCV2 ORF2 and 529 repeat sequence(R epG529)of Toxoplasma gondii(Tox),after optimizing the concentration of primers and probe as well as reaction conditions,a dual TaqMan fluorescence quant itative PCR assay for simultaneous detection o f the two pathogens was established.The method was specific to PCV2 and Tox,without cross-reaction with other pathogens,indicating a good specificity;its limit of detection(LOD)was 1.21×101 cop ies/μL for PCV2 plasmid,and 1.17×101 copies/μL for Tox plasmid,indicating a high sensitivity;intra-and inter-assay coefficients of variation(CVs)were less than 5%,in dic ating a good reproducibility.187 clinical samples of serum and spleen were simultaneously detected by the es tablished method and national standard qPCR,resulting in positive compliance rates of 93.33%and 100%for PCV2 and Tox,respectively.Future high-throughput,rapid and specific detection for PCV2 and Tox was technically supported by the method established by the study.关键词
猪圆环病毒2型/弓形虫/双重荧光定量PCRKey words
PCV2/Toxoplasma gondii/dual fluorescence quantitative PCR分类
农业科技引用本文复制引用
王海燕,仇德洋,蔺雅婷,杜建才,姚方方,吕延飞,刘洋,郑佳..猪圆环病毒2型和弓形虫双重TaqMan荧光定量PCR法的建立及应用[J].中国动物检疫,2025,42(6):92-98,7.基金项目
山东省生猪产业技术体系项目(SDAIT-08-07) （SDAIT-08-07）
