中国兽医科学2025，Vol.55Issue(1)：56-64,9.DOI:10.16656/j.issn.1673-4696.2025.0011
多杀性巴氏杆菌重组酶聚合酶扩增快速诊断方法的建立
Establishment of rapid daignosis method of recombinase polymerase amplification of Pasteurella multocida
摘要
Abstract
The purpose of this study is to establish a sensitive,specific,rapid and visualized clinical diagnostic method based on recombinase polymerase amplification(RPA)and lateral flow im-munochromatography strip(LFD)technology.Specific primers and probes are designed for RPA amplifica-tion and nfo RPA amplification based on species specific Kmt Ⅰ gene of P.multocida.The optimal primers were screened,the reaction system of RPA was optimized,and the optimal reaction conditions were de-termined by agarose gel electrophoresis and lateral flow immunochromatography strip analyzing.The results showed that Kmt Ⅰ could be well detected at 30 ℃ within 20 minutes,and the minimum number of plasmid copies that could be detected was 2.26 × 101 copies/µL.There was no cross-reaction with Es-cherichia coli,Staphylococcus aureus,Salmonella typhimurium,Salmonella cholerae,and Pseudomonas aeruginosa,which showed good specificity.15 clinical samples of Zi Geese were detected RPA basic,RPA-LFD and ordinary PCR,and the results are consistent,with a positive detection rate of 100％and a compliance rate of 100％.In conclusion,RPA basic and RPA-LFD rapid detection methods with high sensi-tivity and specificity,simple operation,low instrument and site requirements for P.multocida detec-tion were established in this study,which provides reliable technical support for the point-of-care testing and epidemiological investigation of established.关键词
多杀性巴氏杆菌/重组酶聚合酶/侧流免疫层析试纸条/快速检测Key words
Pasteurella multitocida/recombinase polymerase amplification/lateral flow strip/rapid detection分类
畜牧业引用本文复制引用
张蕾,杨昊天,姚美玲,南景东,江波涛,史同瑞,黄宣凯,陈亮,冯万宇,苗艳,兰世捷,吴宪,沈思思,张艳,田秋丰..多杀性巴氏杆菌重组酶聚合酶扩增快速诊断方法的建立[J].中国兽医科学,2025,55(1):56-64,9.基金项目
黑龙江省省属科研院所科研业务经费项目(CZKYF2024-1-C016) （CZKYF2024-1-C016）
黑龙江省农业科学院畜牧兽医分院自拟课题(ZNKT-202212) （ZNKT-202212）
黑龙江省农业科技创新跨越工程项目(CX23TS21) （CX23TS21）
齐齐哈尔市科技计划创新激励项目(CNYGG-2023019) （CNYGG-2023019）
