福建农林大学学报（自然科学版）2025，Vol.54Issue(1)：110-116,7.DOI:10.13323/j.cnki.j.fafu(nat.sci.).202405027
兔源支气管败血波氏杆菌荧光定量PCR检测方法的建立
Development of a fluorescence quantitative PCR assay for detection of Bordetella bronchiseptica in rabbits
摘要
Abstract
[Objective]The study aimed to develop an assay for the rapid detection of Bordetella bronchiseptica in rabbits.[Method]A SYBR GreenⅠ-based fluorescence quantitative PCR assay was developed by designing primers and constructing plasmid standard based on the fimX gene of B.bronchiseptica.After evaluating its specificity,sensitiveness and repeatability,the assay was prelimina-rily used for the detection of clinical samples.[Result]The assay had no cross-reaction with other rabbit-sourced bacterial patho-gens,and was only positive for B.bronchiseptica in rabbits,with a detection limit of 13.8 copies·μL-1 of standard plasmid,which was 10-fold higher than that of conventional PCR assay.The variation coefficients for intra-test and inter-test were less than 2% and 3%,respectively.The detection rate of 68 clinical samples via the assay was 39.71%,which was 2.95% higher than that of the con-ventional PCR assay.[Conclusion]The fluorescence quantitative PCR assay based on SYBR GreenⅠprovided an alternative meth-od for the rapid detection of B.bronchiseptica in rabbits with high specificity,high sensitivity and good repeatability,which is helpful for the prevention and control of B.bronchiseptica in rabbits.关键词
兔/兔波氏杆菌病/支气管败血波氏杆菌/荧光定量PCR方法/检出率Key words
rabbit/rabbit bordetellosis/Bordetella bronchiseptica/fluorescence quantitative real-time PCR assay/detection rate分类
农业科技引用本文复制引用
苏进博,王锦祥,殷光文..兔源支气管败血波氏杆菌荧光定量PCR检测方法的建立[J].福建农林大学学报（自然科学版）,2025,54(1):110-116,7.基金项目
福建省科技计划公益类专项(2022R10260012) （2022R10260012）
国家现代农业产业技术体系项目(CARS-43-G-5). （CARS-43-G-5）
