生物技术通报2024，Vol.40Issue(7)：117-124,8.DOI:10.13560/j.cnki.biotech.bull.1985.2023-1107
鸡毒支原体和滑液囊支原体双重LFD-RPA快速检测方法的建立
Establishment of Dual LFD-RPA Rapid Detection Method for Mycoplasma gallisepticum and Mycoplasma synoviae
摘要
Abstract
[Objective]Mycoplasma gallisepticum(MG)and Mycoplasma synoviae(MS)are the two most harmful mycoplasmas to poultry.Early diagnosis of MG and MS was a key to prevent and control avian mycoplasma.Through combining recombinase polymerase amplification(RPA)technology with lateral flow dipstick(LFD),a rapid detection method for MG and MS with convenient operation,rapid response and visual results was established.[Method]Specific primers and probes were designed based on MG mgc2 gene and MS vlhA gene,and a dual LFD-RPA rapid detection method was established by optimizing reaction time,reaction temperature and primer ratio,and its sensitivity,specificity and stability were evaluated.Meanwhile,120 clinical samples were tested.[Result]The amplification was done by dual LFD-RPA detection method at 37℃for 5-10 min,and the optimal primer ratio of RPA-MG-F2/R2 and RPA-MS-F1/R1 was 0.6∶1.4.The minimum detection limits of MG and MS were 3.75 copies/μL and 3.46 copies/μL,respectively.This method and the PCR method recommended by OIE were used to detect 120 samples,and the coincidence rate was 93.3%.[Conclusion]The amplification can be done by the dual LFD-RPA detection method of MG and MS under constant temperature conditions.It has simple operation,rapid response,high sensitivity,strong specificity and good stability,and the results can be observed without using any instrument.It is suitable for the field rapid detection of MG and MS infection alone or mixed infection at the primary level.关键词
鸡毒支原体/滑液囊支原体/重组酶聚合酶扩增技术/测流层析试纸条Key words
Mycoplasma gallisepticum/Mycoplasma synoviae/recombinase polymerase amplification/lateral flow dipstick引用本文复制引用
田兴苗,王健霖,郭磊,司朵朵,龚振兴,李继东..鸡毒支原体和滑液囊支原体双重LFD-RPA快速检测方法的建立[J].生物技术通报,2024,40(7):117-124,8.基金项目
宁夏回族自治区科技创新团队建设项目(2022BSB03107),宁夏银川市校企联合创新专项(2022XQ009),宁夏大学产教融合研究生联合培养示范基地建设项目 （2022BSB03107）
