中国动物传染病学报2024，Vol.32Issue(2)：165-173,9.
鉴别猪圆环病毒2型和3型双重TaqMan MGB探针FQ-PCR检测方法研究
Duplex TaqMan MGB Fluorescence Quantitative Real-Time PCR(FQ-PCR)Assay for the Differential Detection of Porcine Circoviruses 2 and 3
摘要
Abstract
To develop a duplex TaqMan MGB fluorescence quantitative real-time PCR(FQ-PCR)assay for simultaneous and differential detection porcine circovirus 2(PCV2)and PCV3 in the same reaction system,two sets of specific primers for PCV2 and PCV3 and two TaqMan MGB probes were designed for development of a duplex FQ-PCR method according to PCV2 rep gene and PCV3 cap gene sequences.The duplex FQ-PCR method developed was then optimized for its reaction conditions.The results showed that the duplex FQ-PCR specifically detected PCV2 and PCV3 samples and had no cross-reaction with other 8 pathogens and negative controls,The minimum detection limit was 10 copies/μL nucleic acid,which was the same as the single PCV2 or PCV3 FQ-PCR.The coefficients of intra-batch and inter-batch variation were less than 3％,which indicated that the assay had high specificity,sensitivity and repeatability.The interference test demonstrated that the FQ-PCR method had no effect on the detection and accurate quantification of either virus nucleic acid when the initial concentration of the positive plasmids of these viruses were very different.Meanwhile,this method was used to test 16 PCV2,10 PCV3 and 6 PCV2/PCV3 double positive samples and the results were consistent with those of the gene sequencing.The duplex FQ-PCR method developed here had the advantages of high sensitivity,and specificity,simultaneously rapid identification of PCV2 and PCV3,which might be used for the monitoring and surveillance of virus infection.关键词
猪圆环病毒2型/rep基因/猪圆环病毒3型/cap基因/双重TaqMan MGB FQ-PCRKey words
Porcine circovirus 2/rep gene/porcine circovirus 3/cap gene/duplex TaqMan MGB FQ-PCR分类
农业科技引用本文复制引用
赵雪丽,刘影,王东方,闫若潜,王华俊,王淑娟,马震原,谢彩华,柴茂,杨海波,王翠..鉴别猪圆环病毒2型和3型双重TaqMan MGB探针FQ-PCR检测方法研究[J].中国动物传染病学报,2024,32(2):165-173,9.基金项目
2022年度河南省重大科技专项(221100110600) （221100110600）
2023年河南省重点研发专项(231111111300) （231111111300）
河南省现代农业产业技术体系(HARS-22-12-T) （HARS-22-12-T）
