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 1.52A102 plasmid copy number, 100 and 10000 more sensitive than RT-PCR and dot-blot hybridization, respectively.
Highlights
Jembrana Disease Virus (JDV) is a lentivirus associated with an acute disease syndrome on Bali cattle (Bos javanicus) in Indonesia and Australia (Kusumawati et al, 2014a)
We describe the comparison of genomic based-method of reverse transcriptase-polymerase chain reaction (RTPCR), reverse transcriptase-loop mediated isothermal amplification (RT-Loop-Mediated Isothermal Amplification (LAMP)) and dot-blot hybridization to detect JDV genomic material
The aim of this study is to compare the sensitivity among reverse transcriptase-polymerase chain reaction (RT-Polymerase Chain Reaction (PCR)), RT-LAMP and dot-blot hybridization
Summary
Jembrana Disease Virus (JDV) is a lentivirus associated with an acute disease syndrome on Bali cattle (Bos javanicus) in Indonesia and Australia (Kusumawati et al, 2014a). In non-fatal infection, regression of lesions commences about 5 weeks post-infection and the recovered cattle is resistant to further development of clinical disease (Dharma et al, 1991). A recovered cattle will develop a delayed immune respone to the same or different isolates of JDV but still in state of viraemia with no recurrence for at least 2 years after infection (Kusumawati et al, 2014b). The disease have been spread to several islands in Indonesia such as Sumatra and Kalimantan and it is believed occurred via the distribution of carrier cattle (Kusumawati et al, 2014b)
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