Use of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick for an easy and rapid detection of Jembrana disease virus

Kusumawati, Asmarani and Tampubolon, Issabellina Dwades and Hendarta, Narendra Yoga and Salasia, Siti Isrina Oktavia and Wanahari, Tenri Ashari and Mappakaya, Basofi Ashari and Hartati, Sri (2015) Use of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick for an easy and rapid detection of Jembrana disease virus. Use of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick for an easy and rapid detection of Jembrana disease virus, 26. pp. 189-195. ISSN 2347-3584 (Print); 2347-3517 (Electronic)

Text Use of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick for an easy and rapid detection of Jembrana disease virus (2).pdf Download (4MB)

Abstract

Jembrana disease virus (JDV) is a viral pathogen that causes Jembrana disease in Bali cattle (Bos javanicus) with high mortality rate. An easy and rapid diagnostic method is essential for further control this disease. We used a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow dipstick (LFD), based on conserved tm subunit of Jembrana disease virus env gene. The RT-LAMP conditions were optimized by varying the concentration of MgSO4, betaine, dNTP, and temperature as well as the time and duration of reaction. The primers sensitivity for JDV was confirmed. The method was able to detect env-tm gene dilution which contained 2 9 10-15 g of template. Comparatively, the sensitivity of RT-LAMP/LFD was 100-fold more sensitive than reverse transcription-polymerase chain reaction. The primers specificity for JDV was also confirmed using positive andnegative controls. This work also showed that virus detection could be done not only on total RNA extracted from blood but various organs could also be analyzed for the presence of JDV using RT-LAMP/LFD method. The whole process, including the LAMP reaction and the LFD hybridization step only lasts approximately 75 min. Results of analysis can be easily observed with naked eyes without addition of any chemical or further analysis. The combination of RT-LAMP with LFD makes the method a more suitable diagnostic tool in conditions where sophisticated and expensive equipments are not available for field investigations on Jembrana disease in Bali cattle.

Item Type:	Article
Subjects:	Q Science > QR Microbiology > QR355 Virology
Divisions:	Poltekkes Kemenkes Yogyakarta > Jurusan Teknologi Laboratorium Medis > Program Studi DIII Teknologi Laboratorium
Depositing User:	PAK DOSEN 2020
Date Deposited:	09 Nov 2021 21:41
Last Modified:	04 Apr 2023 06:06
URI:	http://eprints.poltekkesjogja.ac.id/id/eprint/7297

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