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【目的】建立一种快速简单检测水霉病病原菌的方法。【方法】针对水霉菌ITS区基因序列设计4条特异性引物,包括两条外引物和两条内引物,优化反应条件,观察检测结果。对该方法的特异性和敏感性进行研究。【结果】建立了环介导等温扩增技术检测水霉菌的方法,确定了其最适反应条件。该方法能够检测到浓度低至103个/mL的水霉菌孢子,其灵敏度是普通PCR方法的100倍。【结论】建立的检测水霉菌的LAMP技术,具有操作简便快速等特点,可用做特异性水霉及其孢子的快速鉴定。
【Objective】 To establish a rapid and simple method for detecting pathogen of watery mildew. 【Method】 Four specific primers were designed according to ITS sequence of water mold fungi, including two external primers and two internal primers. The reaction conditions were optimized and the test results were observed. The specificity and sensitivity of the method were studied. 【Result】 The method of ring-mediated isothermal amplification was established to detect water fungi, and the optimum reaction conditions were determined. This method was able to detect spores of hydromospirillum at concentrations as low as 103 / mL with a sensitivity 100-fold that of the normal PCR method. 【Conclusion】 The established LAMP technology for the detection of fungi in water has the advantages of simple and rapid operation, and can be used as a rapid identification of the specific water mold and its spores.