Establishment of a loop-mediated amplification assay for the specific detection of Saprolegnia and its spores

Abstract:

[Objective] To develop a loop-mediated isothermal amplification (LAMP) method for detection of Saprolegnia. [Methods] Four specific LAMP primers (two inner primers and two outer primers) were designed by targeting ITS gene of Saprolegnia. After optimizing reaction condition, the positive results of LAMP assay were judged through visible green colour. To evaluate the sensitivity and specificity of the assay by detection of Saprolegnia spore samples. [Results] The method of a loop-mediated isothermal amplification (LAMP) for detection of Saprolegnia was established. The sensitivity was 103 spores per mL and was 100 times higher than normal PCR. [Conclusion] LAMP assay is specific and simple to operate without the need of special equipment, our assay presents a promising method for detection of Saprolegnia and its spores.