Development of a Colorimetric Loop-mediated Isothermal Amplification Assay for the Visual Detection of Fusarium oxysporum f.sp. melonis

摘要

Fusarium wilt of melon, Fusarium oxysporum f.sp. melonis is one of the most important diseases, causing tremendous losses in melon growing areas in Iran. There is a real need for a rapid and inexpensive assay to facilitate accurate disease diagnosis and surveillance for better management of Fusarium wilt outbreaks. One of the procedures designed for detection of this disease is loop-mediated isothermal amplification(LAMP)assay;its efficiency has been contrasted with polymerase chain reaction(PCR). The translation elongation factor 1-alpha gene is basically used for designing the LAMP(i.e. F3, B3, FIP, and BIP) together with PCR(F and B). Using hydroxynaphthol blue(HNB) dye, LAMP was placed in a water bath after the optimization was done. The results show LAMP is an advantageous method because it is highly sensitive(100-fold), quite cheap,user-friendly, and safe;in addition, it is performed quickly by visual detection and does not require DNA extraction(in direct-LAMP). The LAMP is believed to be a simple and reliable tool for laboratory purposes because it needs only very basic instruments and the results can be observed and contrasted visually.