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利用玉米细菌性枯萎病菌基因组DNA特有的保守区域设计LAMP(loop-mediated isothermal amplification)引物,通过优化其反应条件,建立了玉米细菌性枯萎病菌的LAMP检测体系。以多种参比菌DNA为模板对LAMP检测体系的特异性进行了验证,利用玉米细菌性枯萎病菌DNA溶液和菌液梯度稀释液对LAMP检测体系的灵敏度进行了验证。在特异性测试中,LAMP检测体系仅对玉米细菌性枯萎病菌进行扩增,对非靶标菌不产生扩增。LAMP检测体系DNA和菌体检测灵敏度分别达到了0.858×10-4ng/uL和45 CFU/mL。为玉米细菌性枯萎病菌的检测提供了一种新的、简便的、快速的检测手段。
LAMP (loop-mediated isothermal amplification) primers were designed based on the conserved regions of the genomic DNA of bacterial wilt pathogens in corn and the LAMP detection system of corn bacterial wilt pathogen was established by optimizing the reaction conditions. The specificity of LAMP detection system was validated by using a variety of reference bacteria DNA as templates. The sensitivity of LAMP detection system was verified by using DNA solution of bacterial wilt pathogen and bacterial liquid dilution. In the specificity test, the LAMP detection system only amplifies the pathogen of Fusarium oxysporum and does not amplify the non-target bacteria. LAMP detection system DNA and cell detection sensitivity reached 0.858 × 10-4ng / uL and 45 CFU / mL. It provides a new, simple and rapid detection method for the detection of corn bacterial wilt pathogen.