Megarani, Dorothea V. and Al-Hussinee, Lowia and Subramaniam, Kuttichantran and Sriwanayos, Preeyanan and Imnoi, Kamonchai and Keleher, Bill and Nicholson, Pamela and Surachetpong, Win and Tattiyapong, Puntanat and Hick, Paul and Gustafson, Lori L. and Waltzek, Thomas B. (2022) Development of a TaqMan quantitative reverse transcription PCR assay to detect tilapia lake virus. DISEASES OF AQUATIC ORGANISMS Dis Aquat Org, 152. pp. 147-158. ISSN 01775103
Development of a TaqMan quantitative reverse.pdf
Restricted to Registered users only
Download (598kB) | Request a copy
Abstract
Tilapia lake virus disease (TiLVD) is an emerging viral disease associated with high morbidity and mortality in cultured tilapia worldwide. In this study, we have developed and
validated a TaqMan quantitative reverse transcription PCR (RT-qPCR) assay for TiLV, targeting a conserved region within segment 10 of the genome. The RT-qPCR assay was efficient (mean ± SD: 96.71 ± 3.20%), sensitive with a limit of detection of 10 RNA viral copies per reaction, and detected TiLV strains from different geographic regions including North America, South America, Africa, and Asia. The intra- and inter-assay variability ranged over 0.18−1.41% and 0.21−2.21%,
respectively. The TaqMan RT-qPCR assay did not cross-react with other RNA viruses of fish, including an orthomyxovirus, a betanodavirus, a picornavirus, and a rhabdovirus. Analysis of 93 proven-positive and 185 proven-negative samples yielded a diagnostic sensitivity of 96.8% and a diagnostic specificity of 100%. The TaqMan RT-qPCR assay also detected TiLV RNA in infected Nile tilapia liver tissue extracts following an experimental challenge study, and it successfully
detected TiLV RNA in SSN-1 (E-11 clone) cell cultures displaying cytopathic effects following their inoculation with TiLV-infected tissue homogenates. Thus, the validated TaqMan RT-qPCR assay should be useful for both research and diagnostic purposes. Additionally, the TiLV qPCR assay returns the clinically relevant viral load of a sample which can assist health professionals in determining the role of TiLV during disease investigations. This RT-qPCR assay could be
integrated into surveillance programs aimed at mitigating the effects of TiLVD on global tilapia production.
Item Type: | Article |
---|---|
Additional Information: | Library Dosen |
Uncontrolled Keywords: | Tilapia; Oreochromis spp.; Tilapia lake virus; Tilapia tilapinevirus; TaqMan; Quantitative PCR; Diagnostic accuracy |
Subjects: | Veterinary Medicine |
Divisions: | Faculty of Veterinary Medicine |
Depositing User: | Erlita Cahyaningtyas Cahyaningtyas |
Date Deposited: | 10 Dec 2024 01:45 |
Last Modified: | 10 Dec 2024 01:45 |
URI: | https://ir.lib.ugm.ac.id/id/eprint/12195 |