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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sg-ntu-dr.10356-1462272023-03-05T16:45:58Z A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages Teoh, Boon-Teong Chin, Kim-Ling Nur-Izyan Samsudin Loong, Shih-Keng Sam, Sing-Sin Tan, Kim-Kee Khor, Chee-Sieng Abd-Jamil, Juraina Zainal, Nurhafiza Wilder-Smith, Annelies Zandi, Keivan Abu Bakar, Sazaly Lee Kong Chian School of Medicine (LKCMedicine) Science::Medicine Infectious Disease Diagnostics 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. Published version 2021-02-02T09:10:22Z 2021-02-02T09:10:22Z 2020 Journal Article Teoh, B.-T., Chin, K.-L., Nur-Izyan Samsudin, Loong, S.-K., Sam, S.-S., Tan, K.-K., . . . Abu Bakar, S. (2020). A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages. BMC Infectious Diseases, 20(1), 947-. doi:10.1186/s12879-020-05585-4 1471-2334 0000-0002-9267-1420 https://hdl.handle.net/10356/146227 10.1186/s12879-020-05585-4 33308203 2-s2.0-85097383069 1 20 en BMC Infectious Diseases © 2020 The Author(s). This 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 giveappropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate ifchanges were made. The images or other third party material in this article are included in the article's Creative Commonslicence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commonslicence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtainpermission directly from the copyright holder. To view a copy of this licence, visithttp://creativecommons.org/licenses/by/4.0/.The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to thedata made available in this article, unless otherwise stated in a credit line to the data. application/pdf |
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Science::Medicine Infectious Disease Diagnostics Teoh, Boon-Teong Chin, Kim-Ling Nur-Izyan Samsudin Loong, Shih-Keng Sam, Sing-Sin Tan, Kim-Kee Khor, Chee-Sieng Abd-Jamil, Juraina Zainal, Nurhafiza Wilder-Smith, Annelies Zandi, Keivan Abu Bakar, Sazaly A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages |
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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. |
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Lee Kong Chian School of Medicine (LKCMedicine) |
author_facet |
Lee Kong Chian School of Medicine (LKCMedicine) Teoh, Boon-Teong Chin, Kim-Ling Nur-Izyan Samsudin Loong, Shih-Keng Sam, Sing-Sin Tan, Kim-Kee Khor, Chee-Sieng Abd-Jamil, Juraina Zainal, Nurhafiza Wilder-Smith, Annelies Zandi, Keivan Abu Bakar, Sazaly |
format |
Article |
author |
Teoh, Boon-Teong Chin, Kim-Ling Nur-Izyan Samsudin Loong, Shih-Keng Sam, Sing-Sin Tan, Kim-Kee Khor, Chee-Sieng Abd-Jamil, Juraina Zainal, Nurhafiza Wilder-Smith, Annelies Zandi, Keivan Abu Bakar, Sazaly |
author_sort |
Teoh, Boon-Teong |
title |
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_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_sort |
reverse transcription loop-mediated isothermal amplification for broad coverage detection of asian and african zika virus lineages |
publishDate |
2021 |
url |
https://hdl.handle.net/10356/146227 |
_version_ |
1759854131514703872 |