The comparison of RT-LAMP, RT-PCR and Dot-Blot hybridization for detection of Jembrana disease virus

Abstract

Jembrana disease virus is a viral pathogen that causes Jembrana disease in Bali cattle (Bos javanicus) with high mortality rate. Infection of Jembrana Disease Virus (JDV) on Bali cattle have caused substantial economic losses for farmers in Indonesia and Australia. In order to control the spread, development of a sensitive detection method is important. In this study, we used three different detection methods based on genomic approach, i.e., reverse transcriptase-polymerase chain reaction (RT-PCR), reverse transcriptase-loop mediated isothermal amplification (RT-LAMP) and dot-blot hybridization to detect JDV. Utilization of pGEX-TM, a recombinant plasmid containing env-tm gene as a positive control showed that RT-LAMP is the most sensitive method compares the two others. It could detect template concentration as low as 10 -6 ng µL -1 or equivalent to 1.52×10 2 plasmid copy number, 100 and 10000 more sensitive than RT-PCR and dot-blot hybridization, respectively.