| Autor: |
Megarani DV; Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610, USA., Al-Hussinee L, Subramaniam K, Sriwanayos P, Imnoi K, Keleher B, Nicholson P, Surachetpong W, Tattiyapong P, Hick P, Gustafson LL, Waltzek TB |
| Jazyk: |
angličtina |
| Zdroj: |
Diseases of aquatic organisms [Dis Aquat Organ] 2022 Dec 22; Vol. 152, pp. 147-158. Date of Electronic Publication: 2022 Dec 22. |
| DOI: |
10.3354/dao03700 |
| Abstrakt: |
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 91 proven-positive and 185 proven-negative samples yielded a diagnostic sensitivity of 96.7% 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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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