动物医学进展2025，Vol.46Issue(10)：43-48,6.
猪鼻支原体RAA快速检测方法的建立和初步应用
Establishment and Preliminary Application of Rapid Detection Method for Mycoplasma porcine rhinoplasma RAA
摘要
Abstract
(Objective)To establish a novel,efficient,and convenient method for detecting Mhr,(Method)This study designed two sets of specific primers based on the P37 gene of Mhr and successfully established a recombinase-mediated isothermal amplification(RAA)detection method for Mhr by optimizing the reac-tion system and conditions.The sensitivity,specificity,and repeatability of the method were evaluated,and the method was applied to preliminary detection of 64 clinical samples suspected or confirmed to be infec-ted with Mhr.(Results)The results showed that the RAA detection method established in this study could complete the amplification of Mhr nucleic acid at 24 ℃ within 20 minutes.The method had no cross-reac-tion with other major respiratory diseases of pigs,such as Mhp,SIV,APP,HPS,and PRRSV,and had good specificity.The minimum detectable limit of the method for Mhr nucleic acid was 1.25×10-4 copies/μL,and the sensitivity was high.By amplifying the positive plasmid with a concentration of 1.25×107 copies/μL three times,clear bands were obtained,indicating good repeatability.The method was applied to detect 64 clinical samples suspected of having respiratory diseases,with a Mhr positive rate of 48.43％(31/64),which was consistent with the detection results obtained by the TaqMan dual-fluorescence PCR method es-tablished in the laboratory for Mhr and porcine respiratory coronavirus.(Conclusion)This study success-fully established a novel RAA method for detecting Mhr,which has the characteristics of specificity,sensi-tivity,rapid reaction,and easy operation,and is suitable for early differential diagnosis in production and laboratory.关键词
猪鼻支原体/重组酶介导等温扩增技术/鉴别诊断检测方法Key words
Mycoplasma suis nasal/Recombinant enzyme mediated isothermal amplification technique/Differential diagnostic test method引用本文复制引用
徐悦,冯明,卢冰霞,许心婷,全琛宇,何颖,胡庭俊,段群棚,赵硕,吴先华,张胜斌,马青兰,张扬祖,金宣讲,覃秀珍..猪鼻支原体RAA快速检测方法的建立和初步应用[J].动物医学进展,2025,46(10):43-48,6.基金项目
国家现代农业产业技术体系广西创新团队建设专项资金资助项目(nycytxgxcxtd-2023-15-02) （nycytxgxcxtd-2023-15-02）
南宁市科技重大专项(20232039) （20232039）
广西基本科研业务费专项(23-4) （23-4）
广西兽医生物技术重点实验室开放研究课题(23-035-32-B-01) （23-035-32-B-01）
广西农业科技自筹经费项目(Z2023033,Z202222,Z2023034) （Z2023033,Z202222,Z2023034）
