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A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection

BACKGROUND: Crimean-Congo Haemorrhagic fever Virus (CCHFV) is a rapidly emerging vector-borne pathogen and the cause of a virulent haemorrhagic fever affecting large parts of Europe, Africa, the Middle East and Asia. METHODOLOGY/PRINCIPLE FINDINGS: An isothermal recombinase polymerase amplification...

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Autores principales: Bonney, Laura C., Watson, Robert J., Afrough, Babak, Mullojonova, Manija, Dzhuraeva, Viktoriya, Tishkova, Farida, Hewson, Roger
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5656326/
https://www.ncbi.nlm.nih.gov/pubmed/29028804
http://dx.doi.org/10.1371/journal.pntd.0006013
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author Bonney, Laura C.
Watson, Robert J.
Afrough, Babak
Mullojonova, Manija
Dzhuraeva, Viktoriya
Tishkova, Farida
Hewson, Roger
author_facet Bonney, Laura C.
Watson, Robert J.
Afrough, Babak
Mullojonova, Manija
Dzhuraeva, Viktoriya
Tishkova, Farida
Hewson, Roger
author_sort Bonney, Laura C.
collection PubMed
description BACKGROUND: Crimean-Congo Haemorrhagic fever Virus (CCHFV) is a rapidly emerging vector-borne pathogen and the cause of a virulent haemorrhagic fever affecting large parts of Europe, Africa, the Middle East and Asia. METHODOLOGY/PRINCIPLE FINDINGS: An isothermal recombinase polymerase amplification (RPA) assay was successfully developed for molecular detection of CCHFV. The assay showed rapid (under 10 minutes) detection of viral extracts/synthetic virus RNA of all 7 S-segment clades of CCHFV, with high target specificity. The assay was shown to tolerate the presence of inhibitors in crude preparations of mock field samples, indicating that this assay may be suitable for use in the field with minimal sample preparation. The CCHFV RPA was successfully used to screen and detect CCHFV positives from a panel of clinical samples from Tajikistan. CONCLUSIONS/SIGNIFICANCE: The assay is a rapid, isothermal, simple-to-perform molecular diagnostic, which can be performed on a light, portable real-time detection device. It is ideally placed therefore for use as a field-diagnostic or in-low resource laboratories, for monitoring of CCHF outbreaks at the point-of-need, such as in remote rural regions in affected countries.
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spelling pubmed-56563262017-11-09 A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection Bonney, Laura C. Watson, Robert J. Afrough, Babak Mullojonova, Manija Dzhuraeva, Viktoriya Tishkova, Farida Hewson, Roger PLoS Negl Trop Dis Research Article BACKGROUND: Crimean-Congo Haemorrhagic fever Virus (CCHFV) is a rapidly emerging vector-borne pathogen and the cause of a virulent haemorrhagic fever affecting large parts of Europe, Africa, the Middle East and Asia. METHODOLOGY/PRINCIPLE FINDINGS: An isothermal recombinase polymerase amplification (RPA) assay was successfully developed for molecular detection of CCHFV. The assay showed rapid (under 10 minutes) detection of viral extracts/synthetic virus RNA of all 7 S-segment clades of CCHFV, with high target specificity. The assay was shown to tolerate the presence of inhibitors in crude preparations of mock field samples, indicating that this assay may be suitable for use in the field with minimal sample preparation. The CCHFV RPA was successfully used to screen and detect CCHFV positives from a panel of clinical samples from Tajikistan. CONCLUSIONS/SIGNIFICANCE: The assay is a rapid, isothermal, simple-to-perform molecular diagnostic, which can be performed on a light, portable real-time detection device. It is ideally placed therefore for use as a field-diagnostic or in-low resource laboratories, for monitoring of CCHF outbreaks at the point-of-need, such as in remote rural regions in affected countries. Public Library of Science 2017-10-13 /pmc/articles/PMC5656326/ /pubmed/29028804 http://dx.doi.org/10.1371/journal.pntd.0006013 Text en © 2017 Bonney et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Bonney, Laura C.
Watson, Robert J.
Afrough, Babak
Mullojonova, Manija
Dzhuraeva, Viktoriya
Tishkova, Farida
Hewson, Roger
A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title_full A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title_fullStr A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title_full_unstemmed A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title_short A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection
title_sort recombinase polymerase amplification assay for rapid detection of crimean-congo haemorrhagic fever virus infection
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5656326/
https://www.ncbi.nlm.nih.gov/pubmed/29028804
http://dx.doi.org/10.1371/journal.pntd.0006013
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