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A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses

OBJECTIVE: The re-emergence and spread of tropical viruses to new areas has raised a wave of concern worldwide. In order to treat patients at an early stage and prevent the diffusion of an outbreak, early diagnosis, and therefore fast and adequate detection, is needed. To this end, a multiplex rever...

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Autores principales: Broeders, Sylvia, Garlant, Linda, Fraiture, Marie-Alice, Vandermassen, Els, Suin, Vanessa, Vanhomwegen, Jessica, Dupont-Rouzeyrol, Myrielle, Rousset, Dominique, Van Gucht, Steven, Roosens, Nancy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Authors. Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7129992/
https://www.ncbi.nlm.nih.gov/pubmed/31884173
http://dx.doi.org/10.1016/j.ijid.2019.12.028
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author Broeders, Sylvia
Garlant, Linda
Fraiture, Marie-Alice
Vandermassen, Els
Suin, Vanessa
Vanhomwegen, Jessica
Dupont-Rouzeyrol, Myrielle
Rousset, Dominique
Van Gucht, Steven
Roosens, Nancy
author_facet Broeders, Sylvia
Garlant, Linda
Fraiture, Marie-Alice
Vandermassen, Els
Suin, Vanessa
Vanhomwegen, Jessica
Dupont-Rouzeyrol, Myrielle
Rousset, Dominique
Van Gucht, Steven
Roosens, Nancy
author_sort Broeders, Sylvia
collection PubMed
description OBJECTIVE: The re-emergence and spread of tropical viruses to new areas has raised a wave of concern worldwide. In order to treat patients at an early stage and prevent the diffusion of an outbreak, early diagnosis, and therefore fast and adequate detection, is needed. To this end, a multiplex reverse transcription real-time polymerase chain reaction TaqMan method was designed to detect Zika (ZIKV) and chikungunya (CHIKV) viruses simultaneously. METHODS: Two methods targeting different genome segments were selected from the literature for each virus. These were adapted for high genome coverage and combined in a four-plex assay that was thoroughly validated in-house. The SCREENED tool was used to evaluate the sequence coverage of the method. RESULTS: The full validation approach showed that the new four-plex method allows the specific and sensitive identification and discrimination of ZIKV and CHIKV in routine samples. The combination of two targets per virus allowing almost 100% coverage of about 500 genomes is shown for the first time. CONCLUSIONS: PCR is a reliable user-friendly technique that can be applied in remote areas. Such multiplex methods enable early and efficient diagnosis, leading to rapid treatment and effective confinement in outbreak cases. They may also serve as an aid for surveillance, and the full validation permits easy method-transfer allowing worldwide harmonization.
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spelling pubmed-71299922020-04-06 A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses Broeders, Sylvia Garlant, Linda Fraiture, Marie-Alice Vandermassen, Els Suin, Vanessa Vanhomwegen, Jessica Dupont-Rouzeyrol, Myrielle Rousset, Dominique Van Gucht, Steven Roosens, Nancy Int J Infect Dis Article OBJECTIVE: The re-emergence and spread of tropical viruses to new areas has raised a wave of concern worldwide. In order to treat patients at an early stage and prevent the diffusion of an outbreak, early diagnosis, and therefore fast and adequate detection, is needed. To this end, a multiplex reverse transcription real-time polymerase chain reaction TaqMan method was designed to detect Zika (ZIKV) and chikungunya (CHIKV) viruses simultaneously. METHODS: Two methods targeting different genome segments were selected from the literature for each virus. These were adapted for high genome coverage and combined in a four-plex assay that was thoroughly validated in-house. The SCREENED tool was used to evaluate the sequence coverage of the method. RESULTS: The full validation approach showed that the new four-plex method allows the specific and sensitive identification and discrimination of ZIKV and CHIKV in routine samples. The combination of two targets per virus allowing almost 100% coverage of about 500 genomes is shown for the first time. CONCLUSIONS: PCR is a reliable user-friendly technique that can be applied in remote areas. Such multiplex methods enable early and efficient diagnosis, leading to rapid treatment and effective confinement in outbreak cases. They may also serve as an aid for surveillance, and the full validation permits easy method-transfer allowing worldwide harmonization. The Authors. Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. 2020-03 2019-12-26 /pmc/articles/PMC7129992/ /pubmed/31884173 http://dx.doi.org/10.1016/j.ijid.2019.12.028 Text en © 2020 The Authors Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Broeders, Sylvia
Garlant, Linda
Fraiture, Marie-Alice
Vandermassen, Els
Suin, Vanessa
Vanhomwegen, Jessica
Dupont-Rouzeyrol, Myrielle
Rousset, Dominique
Van Gucht, Steven
Roosens, Nancy
A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title_full A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title_fullStr A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title_full_unstemmed A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title_short A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
title_sort new multiplex rt-qpcr method for the simultaneous detection and discrimination of zika and chikungunya viruses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7129992/
https://www.ncbi.nlm.nih.gov/pubmed/31884173
http://dx.doi.org/10.1016/j.ijid.2019.12.028
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