DEVELOPMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR RAPID DETECTION OF Colletotrichum gloeosporioides IN MANGO

Anthracnose (Colletotrichum gloeosporioides) is a severe disease from fungi infection causing low productivity of vegetables and fruits. Especially mangoes (Mangifera indica L.), which is a highly economic tropical fruit of Thailand, are exported in the third of the world. Hence, this research aimed to develop the condition of LAMP assay for the detection and identification of Colletotrichum species from mangoes. In the present study, the specificity of a primer set of loop-mediated isothermal amplification (LAMP) targeting glutamine synthase (GLN) and tubulin 2 (TUB2) genes was investigated. The result showed only GLN gene target could successfully detect C. gloeosporioides and presented a high sensitivity at 10 ng/ml, the reaction was performed under a thermal cycler for about 65 °C for 60 minutes. A positive reaction was identified visually from pink color to yellow color and confirmed by 1.5% agarose gel electrophoresis. In conclusion, the LAMP assay for anthracnose in mangoes provided a rapid reaction and no need for complex instruments or systems such as PCR machine. The LAMP assay may therefore be applied in routine diagnosis in the laboratory and for in-field.