A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika vi
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RESEARCH ARTICLE
Open Access
A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages Boon-Teong Teoh1*, Kim-Ling Chin1,2, Nur-Izyan Samsudin1, Shih-Keng Loong1, Sing-Sin Sam1, Kim-Kee Tan1, Chee-Sieng Khor1, Juraina Abd-Jamil1, Nurhafiza Zainal3, Annelies Wilder-Smith4,5, Keivan Zandi3,6 and Sazaly AbuBakar1,3*
Abstract 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 (RTLAMP) 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 RTLAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcriptionpolymerase 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. Keywords: Infectious disease, Diagnostics, RT-LAMP, ZIKV, Mosquito, Vector, Vector-borne
* Correspondence: [email protected]; [email protected] 1 Tropical Infectious Diseases Research and Education Centre (TIDREC), Universiti Malaya, Kuala Lumpur, Malaysia Full list of author information is available at the end of the article © The Author(s). 2020 Open Access 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 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
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