Establishment and Optimization of a Dual Loop-Mediated Isothermal Amplification (LAMP) Rapid Detection System for Sclerotinia sclerotiorum and Alternaria alternata
In order to effectively address the problem of the decline in quality and yield of C. morifolium cv. Chuju caused by the simultaneous infection of two pathogenic fungi, the existing LAMP technology (a detection method that can be achieved without expensive equipment) provides a window period for disease prevention. Developing a LAMP detection method that can simultaneously detect different pathogens and has high sensitivity has become the key to solving this problem. In this study, we first designed specific primers based on the conserved sequences of the two common pathogens of C. morifolium cv. Chuju, namely Sclerotinia sclerotiorum and Alternaria alternata, for LAMP amplification. Subsequently, we gradually optimized each aspect of the LAMP method to enhance the detection efficiency and sensitivity. The LAMP assay showed high specificity for both pathogens and a detection limit of 1.43 × 10？6 ng/μL, which is about 100-fold higher than that of conventional PCR. These improvements aid in the effective monitoring and early diagnosis of disease during the cultivation of C. morifolium cv. Chuju, which will contribute to improved disease management and reduced economic losses.
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