Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR

Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication. Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification. Methodology: A t...

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Main Authors:	M. Saleh Habil, Akrahm, Hamzah, Hairul Aini, Mustafa, Imad Al-Deen, A Talib, Noorlelawati, Abdul Rahman, Siti Nurul Fazlin
Format:	Article
Language:	English
Published:	Kulliyyah of Medicine, International Islamic University Malaysia 2018
Subjects:	QR Microbiology QR355 Virology
Online Access:	http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf

id	iium-66273
recordtype	eprints
spelling	iium-662732019-07-12T03:26:25Z http://irep.iium.edu.my/66273/ Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin QR Microbiology QR355 Virology Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication. Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification. Methodology: A total of 50 serum samples, comprising of 40 HCV-positive and 10 HCV-negative, were included in our study. RNA was extracted, reverse transcribed, and then subjected to real-time PCR amplification. Real-time PCR using EvaGreen dye and primers targeting a 5’UTR was carried out. Reference samples with known viral load were treated similarly to the unknown samples and used to create the standard curves. Results: Our method showed a high level of analytical specificity and accuracy, with a low limit of detection (~2 IU/ml). It yielded repeatable results with less than 4% of intra- assay variation. The assay covered a broad dynamic range of quantification, ranging from 0.34 to 6 log IU/ml. The diagnostic sensitivity, specificity, and accuracy were all 100%, indicating neither false positive nor false negative results were obtained. Conclusion: The developed real time PCR using EvaGreen dye has demonstrated a high analytical and diagnostic performance for HCV quantification, suggesting its potential in clinical diagnosis and management. Kulliyyah of Medicine, International Islamic University Malaysia 2018-08-29 Article PeerReviewed application/pdf en http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf M. Saleh Habil, Akrahm and Hamzah, Hairul Aini and Mustafa, Imad Al-Deen and A Talib, Noorlelawati and Abdul Rahman, Siti Nurul Fazlin (2018) Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR. The International Medical Journal Malaysia, 17 (1). p. 31. E-ISSN 1823-4631 http://iiumedic.net/imjm/v1/download/vol_17_no_3_supp_1/MRS2018-32.pdf
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	QR Microbiology QR355 Virology
spellingShingle	QR Microbiology QR355 Virology M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
description	Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication. Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification. Methodology: A total of 50 serum samples, comprising of 40 HCV-positive and 10 HCV-negative, were included in our study. RNA was extracted, reverse transcribed, and then subjected to real-time PCR amplification. Real-time PCR using EvaGreen dye and primers targeting a 5’UTR was carried out. Reference samples with known viral load were treated similarly to the unknown samples and used to create the standard curves. Results: Our method showed a high level of analytical specificity and accuracy, with a low limit of detection (~2 IU/ml). It yielded repeatable results with less than 4% of intra- assay variation. The assay covered a broad dynamic range of quantification, ranging from 0.34 to 6 log IU/ml. The diagnostic sensitivity, specificity, and accuracy were all 100%, indicating neither false positive nor false negative results were obtained. Conclusion: The developed real time PCR using EvaGreen dye has demonstrated a high analytical and diagnostic performance for HCV quantification, suggesting its potential in clinical diagnosis and management.
format	Article
author	M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin
author_facet	M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin
author_sort	M. Saleh Habil, Akrahm
title	Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
title_short	Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
title_full	Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
title_fullStr	Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
title_full_unstemmed	Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
title_sort	establishment of simultaneous detection and quantitation of hcv-rna by third generation intercalating dye real-time pcr
publisher	Kulliyyah of Medicine, International Islamic University Malaysia
publishDate	2018
url	http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf
first_indexed	2023-09-18T21:34:04Z
last_indexed	2023-09-18T21:34:04Z
_version_	1777412703256576000

Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR

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