Novel duplex RT-qPCR for animal rabies surveillance.

Autor: Minozzo GA; Molecular Biology Section, Central Laboratory of Parana State, São José dos Pinhais, Paraná Sate, Brazil.; Graduate College of Cellular and Molecular Biology, Federal University of Paraná (UFPR), Curitiba, Paraná State, Brazil., Corona TF; Molecular Biology Section, Central Laboratory of Parana State, São José dos Pinhais, Paraná Sate, Brazil., da Cruz ECR; Molecular Biology Section, Central Laboratory of Parana State, São José dos Pinhais, Paraná Sate, Brazil., de Castro WAC; Latin-American Institute of Life and Nature Sciences, Federal University for Latin American Integration (UNILA), Foz do Iguaçu, PR, Brazil., Kmetiuk LB; Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, Indiana, 47907-2027, USA., Dos Santos AP; Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, Indiana, 47907-2027, USA., Biondo AW; Department of Veterinary Medicine, Federal University of Paraná State, Curitiba, Paraná State, Brazil., Riediger IN; Molecular Biology Section, Central Laboratory of Parana State, São José dos Pinhais, Paraná Sate, Brazil.
Jazyk: angličtina
Zdroj: Transboundary and emerging diseases [Transbound Emerg Dis] 2022 Sep; Vol. 69 (5), pp. e2261-e2267. Date of Electronic Publication: 2022 Apr 28.
DOI: 10.1111/tbed.14565
Abstrakt: Rabies is a lethal zoonosis affecting mammals worldwide. Diagnosis of rabies follows international standard protocols, primarily relying on direct immunofluorescence (DI) followed by mouse inoculation test (MIT). WHO recommends molecular biology techniques such as RT-qPCR for replacing MIT to diagnose rabies in animal samples. Recently, a real-time PCR protocol that detects all rabies virus variants identified worldwide was validated. This assay is a pan-Lyssavirus TaqMan quantitative RT-PCR called LN34. A modified LN34 assay protocol was tested at the Paraná State Reference Laboratory (Lacen/PR) using animal samples previously tested by DI and MIT, the gold standard (GS). This method has been changed to a RT-qPCR duplex format to better fit the diagnostic routine. The new assay was called duplex LN34 and β-actin RT-qPCR. All the 88 samples evaluated using the GS test, modified pan-Lyssavirus TaqMan RT-qPCR and duplex LN34 and β-actin RT-qPCR showed 100% agreement with each other. This novel duplex RT-qPCR protocol has shown adequate diagnostic performance and may be used in research and surveillance purposes, replacing the standard MIT and ending mice use for rabies diagnosis.
(© 2022 Wiley-VCH GmbH.)
Databáze: MEDLINE
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