Research Article Open Access

The Comparison of RT-LAMP, RT-PCR and Dot-Blot Hybridization for Detection of Jembrana Disease Virus

Asmarani Kusumawati1, Tenri A. Wanahari2, Issabellina D. Tampubolon1 and Basofi A. Mappakaya2
  • 1 Gadjah Mada University, Indonesia
  • 2 Sebelas Maret University, Indonesia

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 1.52×102 plasmid copy number, 100 and 10000 more sensitive than RT-PCR and dot-blot hybridization, respectively.

American Journal of Biochemistry and Biotechnology
Volume 11 No. 2, 2015, 114-118

DOI: https://doi.org/10.3844/ajbbsp.2015.114.118

Submitted On: 19 May 2015 Published On: 26 June 2015

How to Cite: Kusumawati, A., Wanahari, T. A., Tampubolon, I. D. & Mappakaya, B. A. (2015). The Comparison of RT-LAMP, RT-PCR and Dot-Blot Hybridization for Detection of Jembrana Disease Virus. American Journal of Biochemistry and Biotechnology, 11(2), 114-118. https://doi.org/10.3844/ajbbsp.2015.114.118

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Keywords

  • Jembrana Disease
  • RT-PCR
  • RT-LAMP
  • dot-blot hybridization
  • Comparative Analysis