可可花瘿病菌LAMP检测方法的建立OA

[Aim]This study aims to develop a rapid,sensitive,convenient,and visual loop-mediated isothermal amplification(LAMP)detection method for Albonectria rigidiuscula,providing technical support for the rapid detection and accurate management of this invasive pest.[Method]LAMP primers were designed based on the EF-1α and Tub sequences of A.rigidiuscula,and a set of effective primers was screened.The reaction temperature was optimized,and the specificity and sensitivity of the method were evaluated.Additionally,the host of A.rigidiuscula was detected.[Result]The visual LAMP detection method established in this study achieved the detection of A.rigidiuscula within 1 h.No amplification band was produced for the 19 other pathogenic strains tested,and only A.rigidiuscula exhibited amplification,which demonstrated the excellent specificity of the established method.The optimal detection temperature was 65℃,and the detection sensitivity reached 21.84×10-4 ng•μL-1.In practical applications,the method achieved an accuracy rate of 100％,and the test results could be intuitively interpreted.[Conclusion]The LAMP method developed in this study for the detection of A.rigidiuscula features a short reaction time,strong specificity,high sensitivity,simple operation,and visual detection results.It is suitable for port quarantine and field detection of A.rigidiuscula.