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A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages

BACKGROUND: Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in res...

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Autores principales: Teoh, Boon-Teong, Chin, Kim-Ling, Samsudin, Nur-Izyan, Loong, Shih-Keng, Sam, Sing-Sin, Tan, Kim-Kee, Khor, Chee-Sieng, Abd-Jamil, Juraina, Zainal, Nurhafiza, Wilder-Smith, Annelies, Zandi, Keivan, AbuBakar, Sazaly
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731766/
https://www.ncbi.nlm.nih.gov/pubmed/33308203
http://dx.doi.org/10.1186/s12879-020-05585-4
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author Teoh, Boon-Teong
Chin, Kim-Ling
Samsudin, Nur-Izyan
Loong, Shih-Keng
Sam, Sing-Sin
Tan, Kim-Kee
Khor, Chee-Sieng
Abd-Jamil, Juraina
Zainal, Nurhafiza
Wilder-Smith, Annelies
Zandi, Keivan
AbuBakar, Sazaly
author_facet Teoh, Boon-Teong
Chin, Kim-Ling
Samsudin, Nur-Izyan
Loong, Shih-Keng
Sam, Sing-Sin
Tan, Kim-Kee
Khor, Chee-Sieng
Abd-Jamil, Juraina
Zainal, Nurhafiza
Wilder-Smith, Annelies
Zandi, Keivan
AbuBakar, Sazaly
author_sort Teoh, Boon-Teong
collection PubMed
description BACKGROUND: Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in resource-deficient settings, is highly required. METHODS: In the present study, a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of both the Asian and African-lineage ZIKV. The detection limit, strain coverage and cross-reactivity of the ZIKV RT-LAMP assay was evaluated. The sensitivity and specificity of the RT-LAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used as the reference assay. RESULTS: The detection limit of the RT-LAMP assay was 3.73 ZIKV RNA copies (probit analysis, P ≤ 0.05). The RT-LAMP assay detected the ZIKV genomes of both the Asian and African lineages without cross-reacting with other arthropod-borne viruses. The sensitivity and specificity of the RT-LAMP assay were 90% (95% CI = 59.6–98.2) and 100% (95% CI = 78.5–100.0), respectively. The RT-LAMP assay detected ZIKV genome in 9 of 24 (37.5%) of the simulated clinical samples compared to 10 of 24 (41.7%) by qRT-PCR assay with a high level of concordance (κ = 0.913, P < 0.001). CONCLUSION: The RT-LAMP assay is applicable for the broad coverage detection of both the Asian and African ZIKV strains in resource-deficient settings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-020-05585-4.
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spelling pubmed-77317662020-12-15 A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages Teoh, Boon-Teong Chin, Kim-Ling Samsudin, Nur-Izyan Loong, Shih-Keng Sam, Sing-Sin Tan, Kim-Kee Khor, Chee-Sieng Abd-Jamil, Juraina Zainal, Nurhafiza Wilder-Smith, Annelies Zandi, Keivan AbuBakar, Sazaly BMC Infect Dis Research Article BACKGROUND: Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in resource-deficient settings, is highly required. METHODS: In the present study, a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of both the Asian and African-lineage ZIKV. The detection limit, strain coverage and cross-reactivity of the ZIKV RT-LAMP assay was evaluated. The sensitivity and specificity of the RT-LAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used as the reference assay. RESULTS: The detection limit of the RT-LAMP assay was 3.73 ZIKV RNA copies (probit analysis, P ≤ 0.05). The RT-LAMP assay detected the ZIKV genomes of both the Asian and African lineages without cross-reacting with other arthropod-borne viruses. The sensitivity and specificity of the RT-LAMP assay were 90% (95% CI = 59.6–98.2) and 100% (95% CI = 78.5–100.0), respectively. The RT-LAMP assay detected ZIKV genome in 9 of 24 (37.5%) of the simulated clinical samples compared to 10 of 24 (41.7%) by qRT-PCR assay with a high level of concordance (κ = 0.913, P < 0.001). CONCLUSION: The RT-LAMP assay is applicable for the broad coverage detection of both the Asian and African ZIKV strains in resource-deficient settings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-020-05585-4. BioMed Central 2020-12-11 /pmc/articles/PMC7731766/ /pubmed/33308203 http://dx.doi.org/10.1186/s12879-020-05585-4 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Teoh, Boon-Teong
Chin, Kim-Ling
Samsudin, Nur-Izyan
Loong, Shih-Keng
Sam, Sing-Sin
Tan, Kim-Kee
Khor, Chee-Sieng
Abd-Jamil, Juraina
Zainal, Nurhafiza
Wilder-Smith, Annelies
Zandi, Keivan
AbuBakar, Sazaly
A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title_full A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title_fullStr A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title_full_unstemmed A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title_short A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages
title_sort reverse transcription loop-mediated isothermal amplification for broad coverage detection of asian and african zika virus lineages
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731766/
https://www.ncbi.nlm.nih.gov/pubmed/33308203
http://dx.doi.org/10.1186/s12879-020-05585-4
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