Tracking and identification of Alfalfa Mosaic Virus (AMV) by Loop mediated isothermal amplification assay

Alfalfa mosaic virus (AMV) causing a disease in Alfalfa (Medicago sativa) crop in Iran has been identified on the basis of determination of symptom expression and morphological properties. There are several techniques to detect the virus including serological test and molecular methods. Reverse transcription Loop-mediated isothermal amplification (RT-LAMP) assay is a novel technique for amplifying RNA under constant temperature, with high specificity, sensitivity, rapidity and efficiency that is used in this study for detection of Alfalfa mosaic virus. Leaf samples (100 samples) with symptoms similar to AMV were collected from Hamedan and Kurdestan provinces and were subjected to a serological test. All four RT-LAMP reaction primers (i.e. F3, B3, FIP and BIP) together with RT-PCR reaction primers were designed on the basis of the highly conserved sequence of coat protein (CP) gene. Total RNA was extracted and molecular reactions were carried out and finally was detected six positive sampled The advantages of this new method in compare to other methods include the high specificity, high sensitivity, high rapidity, high efficiency, safety, fascinatingly, no requirement of expensive and tools for amplification, no post-amplification treatment of the amplicons, visual detection and user friendly.