A recombinase polymerase amplification assay for rapid detection of rabies virus

Abstract Rabies is a generally fatal encephalitis caused by a negative-sense single-stranded RNA lyssavirus transmitted to humans mainly from dog bite. Despite the recommendation by WHO and OIE to use the direct immunofluorescence test as standard method, molecular diagnostic assays like reverse tra...

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Autores principales:	Martin Faye, Ahmed Abd El Wahed, Oumar Faye, Jonas Kissenkötter, Bernd Hoffmann, Amadou Alpha Sall, Ousmane Faye
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5b7dcb9c932146a6932770deb86cbee8
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spelling	oai:doaj.org-article:5b7dcb9c932146a6932770deb86cbee82021-12-02T14:06:32ZA recombinase polymerase amplification assay for rapid detection of rabies virus10.1038/s41598-021-82479-82045-2322https://doaj.org/article/5b7dcb9c932146a6932770deb86cbee82021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82479-8https://doaj.org/toc/2045-2322Abstract Rabies is a generally fatal encephalitis caused by a negative-sense single-stranded RNA lyssavirus transmitted to humans mainly from dog bite. Despite the recommendation by WHO and OIE to use the direct immunofluorescence test as standard method, molecular diagnostic assays like reverse transcription quantitative polymerase chain reaction (RT-qPCR) are increasing as a confirmatory method. However, both technologies are inaccessible in resource-limited settings. Moreover, the available point-of-need molecular assay is of poor detection limit for African strains. Herein, we developed a reverse transcription recombinase polymerase amplification (RT-RPA) assay as potential point-of-need diagnostic tool for rapid detection of various strains of rabies virus including locally isolated African strains. The sensitivity and specificity of the method was evaluated using a molecular RNA standard and different Rabies-related viruses belonging to the Rhabdoviridea family, respectively. The RABV-RPA performances were evaluated on isolates representative of the existing diversity and viral dilutions spiked in non-neural clinical specimen. The results were compared with RT-qPCR as a gold standard. The RABV-RPA detected down to 4 RNA molecules per reaction in 95% of the cases in less than 10 min. The RABV-RPA assay is highly specific as various RABV isolates were identified, but no amplification was observed for other member of the Rhabdoviridea family. The sample background did not affect the performance of the RABV-RPA as down to 11 RNA molecules were identified, which is similar to the RT-qPCR results. Our developed assay is suitable for use in low-resource settings as a promising alternative tool for ante-mortem rabies diagnosis in humans for facilitating timely control decisions.Martin FayeAhmed Abd El WahedOumar FayeJonas KissenkötterBernd HoffmannAmadou Alpha SallOusmane FayeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Martin Faye Ahmed Abd El Wahed Oumar Faye Jonas Kissenkötter Bernd Hoffmann Amadou Alpha Sall Ousmane Faye A recombinase polymerase amplification assay for rapid detection of rabies virus
description	Abstract Rabies is a generally fatal encephalitis caused by a negative-sense single-stranded RNA lyssavirus transmitted to humans mainly from dog bite. Despite the recommendation by WHO and OIE to use the direct immunofluorescence test as standard method, molecular diagnostic assays like reverse transcription quantitative polymerase chain reaction (RT-qPCR) are increasing as a confirmatory method. However, both technologies are inaccessible in resource-limited settings. Moreover, the available point-of-need molecular assay is of poor detection limit for African strains. Herein, we developed a reverse transcription recombinase polymerase amplification (RT-RPA) assay as potential point-of-need diagnostic tool for rapid detection of various strains of rabies virus including locally isolated African strains. The sensitivity and specificity of the method was evaluated using a molecular RNA standard and different Rabies-related viruses belonging to the Rhabdoviridea family, respectively. The RABV-RPA performances were evaluated on isolates representative of the existing diversity and viral dilutions spiked in non-neural clinical specimen. The results were compared with RT-qPCR as a gold standard. The RABV-RPA detected down to 4 RNA molecules per reaction in 95% of the cases in less than 10 min. The RABV-RPA assay is highly specific as various RABV isolates were identified, but no amplification was observed for other member of the Rhabdoviridea family. The sample background did not affect the performance of the RABV-RPA as down to 11 RNA molecules were identified, which is similar to the RT-qPCR results. Our developed assay is suitable for use in low-resource settings as a promising alternative tool for ante-mortem rabies diagnosis in humans for facilitating timely control decisions.
format	article
author	Martin Faye Ahmed Abd El Wahed Oumar Faye Jonas Kissenkötter Bernd Hoffmann Amadou Alpha Sall Ousmane Faye
author_facet	Martin Faye Ahmed Abd El Wahed Oumar Faye Jonas Kissenkötter Bernd Hoffmann Amadou Alpha Sall Ousmane Faye
author_sort	Martin Faye
title	A recombinase polymerase amplification assay for rapid detection of rabies virus
title_short	A recombinase polymerase amplification assay for rapid detection of rabies virus
title_full	A recombinase polymerase amplification assay for rapid detection of rabies virus
title_fullStr	A recombinase polymerase amplification assay for rapid detection of rabies virus
title_full_unstemmed	A recombinase polymerase amplification assay for rapid detection of rabies virus
title_sort	recombinase polymerase amplification assay for rapid detection of rabies virus
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/5b7dcb9c932146a6932770deb86cbee8
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A recombinase polymerase amplification assay for rapid detection of rabies virus

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