Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus

Due to the general symptoms presented by the Chikungunya virus (CHIKV)-infected patients, a laboratory test is needed to differentiate CHIKV from other viral infections. The reverse transcription-quantitative real-time PCR (RT-qPCR) is a rapid and sensitive diagnostic tool, and several assays hav...

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Main Authors: Anna, Andrew, Citartan, Marimuthu, Wong, Kiing Aik, Tang, Thean Hock, Sum, Magdline Sia Henry, Ch'ng, Ewe Seng
Format: Article
Language:English
Published: 2023
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Online Access:http://ir.unimas.my/id/eprint/42216/1/Anna%20Andrew.pdf
http://ir.unimas.my/id/eprint/42216/
https://journals.asm.org/doi/epub/10.1128/spectrum.00088-23
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spelling my.unimas.ir.422162023-07-11T01:44:13Z http://ir.unimas.my/id/eprint/42216/ Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus Anna, Andrew Citartan, Marimuthu Wong, Kiing Aik Tang, Thean Hock Sum, Magdline Sia Henry Ch'ng, Ewe Seng QR Microbiology QR355 Virology Due to the general symptoms presented by the Chikungunya virus (CHIKV)-infected patients, a laboratory test is needed to differentiate CHIKV from other viral infections. The reverse transcription-quantitative real-time PCR (RT-qPCR) is a rapid and sensitive diagnostic tool, and several assays have been developed for detecting and quantifying CHIKV. Since real-time amplification efficiency varies within and between laboratories, an assay must be validated before being used on patient samples. In this study, the diagnostic performance of a TaqMan RT-qPCR assay was evaluated using synthetic RNA and archived patient samples. The cutoff quantification cycle (Cq) value for the assay was determined by experimental evidence. We found the in-house assay was highly sensitive, with a detection limit of 3.95 RNA copies/reaction. The analytical specificity of the assay was 100%. The analytical cutoff Cq value was 37, corresponding to the mean Cq value of the detection limit. Using archived samples characterized previously, the sensitivity and specificity of the assay were 76% and 100%, respectively. The inhouse assay was also compared with a commercial assay, and we found that the inhouse assay had higher sensitivity. Although further evaluation with prospective patient samples is needed in the future, this validated RT-qPCR was sensitive and specific, which shows its potential to detect CHIKV in clinical samples 2023 Article PeerReviewed text en http://ir.unimas.my/id/eprint/42216/1/Anna%20Andrew.pdf Anna, Andrew and Citartan, Marimuthu and Wong, Kiing Aik and Tang, Thean Hock and Sum, Magdline Sia Henry and Ch'ng, Ewe Seng (2023) Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus. Microbiology Spectrum. pp. 1-9. ISSN 2165-0497 https://journals.asm.org/doi/epub/10.1128/spectrum.00088-23 10.1128/spectrum.00088-23
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic QR Microbiology
QR355 Virology
spellingShingle QR Microbiology
QR355 Virology
Anna, Andrew
Citartan, Marimuthu
Wong, Kiing Aik
Tang, Thean Hock
Sum, Magdline Sia Henry
Ch'ng, Ewe Seng
Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
description Due to the general symptoms presented by the Chikungunya virus (CHIKV)-infected patients, a laboratory test is needed to differentiate CHIKV from other viral infections. The reverse transcription-quantitative real-time PCR (RT-qPCR) is a rapid and sensitive diagnostic tool, and several assays have been developed for detecting and quantifying CHIKV. Since real-time amplification efficiency varies within and between laboratories, an assay must be validated before being used on patient samples. In this study, the diagnostic performance of a TaqMan RT-qPCR assay was evaluated using synthetic RNA and archived patient samples. The cutoff quantification cycle (Cq) value for the assay was determined by experimental evidence. We found the in-house assay was highly sensitive, with a detection limit of 3.95 RNA copies/reaction. The analytical specificity of the assay was 100%. The analytical cutoff Cq value was 37, corresponding to the mean Cq value of the detection limit. Using archived samples characterized previously, the sensitivity and specificity of the assay were 76% and 100%, respectively. The inhouse assay was also compared with a commercial assay, and we found that the inhouse assay had higher sensitivity. Although further evaluation with prospective patient samples is needed in the future, this validated RT-qPCR was sensitive and specific, which shows its potential to detect CHIKV in clinical samples
format Article
author Anna, Andrew
Citartan, Marimuthu
Wong, Kiing Aik
Tang, Thean Hock
Sum, Magdline Sia Henry
Ch'ng, Ewe Seng
author_facet Anna, Andrew
Citartan, Marimuthu
Wong, Kiing Aik
Tang, Thean Hock
Sum, Magdline Sia Henry
Ch'ng, Ewe Seng
author_sort Anna, Andrew
title Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
title_short Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
title_full Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
title_fullStr Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
title_full_unstemmed Analytical and Clinical Evaluation of a TaqMan Real-Time PCR Assay for the Detection of Chikungunya Virus
title_sort analytical and clinical evaluation of a taqman real-time pcr assay for the detection of chikungunya virus
publishDate 2023
url http://ir.unimas.my/id/eprint/42216/1/Anna%20Andrew.pdf
http://ir.unimas.my/id/eprint/42216/
https://journals.asm.org/doi/epub/10.1128/spectrum.00088-23
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score 13.159267