Development of a TaqMan minor groove binding probe-based quantitative reverse transcription polymerase chain reaction for the detection and quantification of Zika virus

Chin, Kim Ling and Teoh, Boon Teong and Sam, Sing Sin and Loong, Shih Keng and Tan, K. K. and Azizan, N. S. and Lim, Y. K. and Khor, Chee Sieng and Nor'e, S. S. and Abd-Jamil, Juraina and Abu Bakar, Sazaly (2022) Development of a TaqMan minor groove binding probe-based quantitative reverse transcription polymerase chain reaction for the detection and quantification of Zika virus. TROPICAL BIOMEDICINE, 39 (4). pp. 518-523. ISSN 0127-5720, DOI https://doi.org/10.47665/tb.39.4.005.

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Official URL: https://doi.org/10.47665/tb.39.4.005

Abstract

Zika virus (ZIKV) infection has emerged as a global health concern following epidemic outbreaks of severe neurological disorders reported in Pacific and Americas since 2016. Therefore, a rapid, sensitive and specific diagnostic test for ZIKV infection is critical for the appropriate patient management and the control of disease spread. A TaqMan minor groove binding (MGB) probe-based quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed based on the conserved sequence regions of 463 ZIKV NS2B genes. The designed ZIKV qRT-PCR assay was evaluated for its detection limit, strain coverage and cross-reactivity. We further assessed the clinical applicability of qRT-PCR assay for ZIKV RNA detection using a total 18 simulated clinical specimens. The detection limit of the qRT-PCR assay was 11.276 ZIKV RNA copies at the 95% probability level (probit analysis, p< 0.05). Both Asian and African ZIKV strains were detected by the qRT-PCR assay without cross-reacting with DENV-1, DENV-2, DENV-3, DENV-4, CHIKV, JEV, LGTV, GETV and SINV. The qRT-PCR assay demonstrated a perfect agreement (k = 1.000, P < 0.001) with the reference assay; the sensitivity and specificity of the qRT-PCR assay were 100% (95% CI= 79.6-100) and 100% (95% CI= 43.9-100) respectively. The qRT-PCR assay developed in this study is a useful diagnostic tool for the broad coverage detection and quantification of both the Asian and African ZIKV strains.

Item Type: Article
Funders: Ministry of Education Malaysia for niche area research under the Higher Institution Centre of Excellence (HICoE) program [MO002-2019], Long Term Research Grant Scheme [LRGS MRUN/F1/01/2018]
Uncontrolled Keywords: Infectious disease; vector-borne; mosquito; MGB probe; diagnostics
Subjects: Q Science > QR Microbiology
R Medicine > RB Pathology
Divisions: Faculty of Medicine > Medical Microbiology Department
Institute of Advanced Studies
Deputy Vice Chancellor (Research & Innovation) Office > Tropical Infectious Diseases Research and Education Centre
Depositing User: Ms Koh Ai Peng
Date Deposited: 01 Nov 2024 08:15
Last Modified: 01 Nov 2024 08:15
URI: http://eprints.um.edu.my/id/eprint/46129

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