One-step real-time loop-mediated isothermal amplification (RT-LAMP): evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes

Authors : Sidra MARYAM, Tahir RASHEED, Asma LATIF, Rabaab ZAHRA, Aamer Bin ZAHUR, Aitezaz AHSAN, Muhammad AFZAL, Umer FAROOQ

Pages : 435-443

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Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :Availability of a rapid and sensitive diagnostic technique is key in successful prevention and control of infectious diseases like foot-and-mouth disease insert ignore into journalissuearticles values(FMD);. Existing conventional diagnostic tests for FMD are laborious and time-consuming with low sensitivity and specificity. Molecular-based techniques are costly and difficult, involving refined apparatus like a thermal cycler. In the present study, the technique of real-time loop-mediated isothermal amplification insert ignore into journalissuearticles values(RT-LAMP); was standardized for diagnosing FMDV and its serotypes, evaluated using field samples, and compared with the existing real-time PCR in Pakistan. RT-LAMP amplified the target 3D gene using specific primers at 65 °C for 60 min and the VP1 gene using serotype specific primers at 63 °C for 60 min. A total of 38 samples out of 50 were positive by RT-LAMP and identified serotypes were A insert ignore into journalissuearticles values(n = 15);, O insert ignore into journalissuearticles values(n = 15);, and Asia-1 insert ignore into journalissuearticles values(n = 8);. The efficiency of RT-LAMP in this study was highest for serotype Asia-1 insert ignore into journalissuearticles values(80.9%); followed by serotype A insert ignore into journalissuearticles values(73.4%); and O insert ignore into journalissuearticles values(62.35);.
Keywords : Reverse transcriptase loop mediated isothermal amplification, foot and mouth disease, real time polymerase chain reaction