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

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

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Abstract

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.

Item Type:	Article (Journal)
Additional Information:	5342/66273
Uncontrolled Keywords:	Hepatitis C, EvaGreen dye, Real-time PCR, viral load
Subjects:	Q Science > QR Microbiology Q Science > QR Microbiology > QR355 Virology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Medicine Kulliyyah of Medicine > Department of Basic Medical Kulliyyah of Medicine > Department of Pathology & Lab Medicine
Depositing User:	Dr Hairul Aini Hamzah
Date Deposited:	10 Apr 2019 16:05
Last Modified:	12 Jul 2019 11:26
URI:	http://irep.iium.edu.my/id/eprint/66273

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