Establishment and Preliminary Application of Recombinase-mediated Amplification-sidestream Detection for Detection of Mycobacterium paratuberculosis
To establish a rapid and accurate method for detection of Mycobacterium avium subsp.paratuberculosis(MAP),primers and probes were designed according to the specific multicopy insert element IS900 of MAP.A recombinase-aid amplification combined with lateral flow dipstick(RAA-LFD)method was established for visual detection of MAP in the field within 30 min at 35 ℃ after being optimized.The assay was specific with the target sequence of MAP with a detection limit up to 10 fg·μL-1.One hundred and forty-nine stool samples were tested by this method,showing 100％sensitivity with the PCR.In short,the RAA-LFD assay developed in this study was a rapid,reliable method which can be broadly used.
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