Real-Time loop-mediated isothermal amplification assay for rapid detection of Banana bunchy top virus in North-east India

Aim: The study aims to standardize a Real-Time LAMP assay for effective, highly sensitive, and rapid detection of BBTV in North-east India.Methodology: symptoms were collected from Assam, India and subjected to conventional PCR for confirmation. Six sets of BBTV LAMP primers were designed and the PCR positive samples were subjected to Real -Time LAMP assay for detection of BBTV. Finally, a sensitivity test of BBTV LAMP assay and comparison of BBTV LAMP assay with conventional PCR was done using seven 10-fold dilutions of total genomic DNA of leaf samples with the highest dilution starting from 100 ng mu l .-1 Forty samples of banana showing BBTV likeResults: Initially a total of twenty six out of forty banana samples were tested positive for BBTV with conventional PCR method. The Real -Time LAMP assay for BBTV detection resulted in typical sigmoidal amplification curves with the peak values ranging between 8.00 to 12.15 min and annealing derivatives ranging between 83.3 C to o84.3 C in the tested samples. Sensitivity testing and comparison of o BBTV Real-Time LAMP assay with conventional PCR revealed that the BBTV LAMP assay could efficiently detect up to 0.0001ng mu l of-1 total DNA against 0.01ng mu l in conventional PCR.-1Interpretation: effective diagnosis of BBTV using Real-Time LAMP method. This method can be preferred over conventional diagnostic techniques like PCR or ELISA for rapid large scale detection of BBTV in banana plants in North-east India. The findings highlight rapid, sensitive, accurate and