Review
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 28, 2016; 22(20): 4824-4834
Published online May 28, 2016. doi: 10.3748/wjg.v22.i20.4824
Multiplex qPCR for serodetection and serotyping of hepatitis viruses: A brief review
Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari
Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India
Author contributions: Gupta P collected the information from published literature; Mankotia DS and Ansari MA categorized it under different sub-headings and prepared the manuscript; Irshad M edited the manuscript and corrected/modified the language.
Conflict-of-interest statement: There is no conflict of interest among the authors of this study.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Dr. Mohammad Irshad, Professor, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. drirshad54@yahoo.com
Telephone: +91-11-26594981 Fax: +91-11-26588663
Received: February 9, 2016
Peer-review started: February 9, 2016
First decision: March 7, 2016
Revised: March 9, 2016
Accepted: March 30, 2016
Article in press: March 30, 2016
Published online: May 28, 2016
Processing time: 100 Days and 0 Hours
Abstract

The present review describes the current status of multiplex quantitative real time polymerase chain reaction (qPCR) assays developed and used globally for detection and subtyping of hepatitis viruses in body fluids. Several studies have reported the use of multiplex qPCR for the detection of hepatitis viruses, including hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), and hepatitis E virus (HEV). In addition, multiplex qPCR has also been developed for genotyping HBV, HCV, and HEV subtypes. Although a single step multiplex qPCR assay for all six hepatitis viruses, i.e., A to G viruses, is not yet reported, it may be available in the near future as the technologies continue to advance. All studies use a conserved region of the viral genome as the basis of amplification and hydrolysis probes as the preferred chemistries for improved detection. Based on a standard plot prepared using varying concentrations of template and the observed threshold cycle value, it is possible to determine the linear dynamic range and to calculate an exact copy number of virus in the specimen. Advantages of multiplex qPCR assay over singleplex or other molecular techniques in samples from patients with co-infection include fast results, low cost, and a single step investigation process.

Keywords: Co-infection; Viral genome; Quantitative real-time polymerase chain reaction; Genotyping techniques; Serotyping; Hepatitis viruses

Core tip: The present review describes the worldwide application and the significance of multiplex quantitative real time polymerase chain reaction (qPCR) for simultaneous detection of hepatitis viruses and their subtypes in serum. The published literature has demonstrated that the multiplex qPCR assay is a fast, easy, cost-effective, and sensitive technique for the early diagnosis of hepatitis co-infections. Use of this technique, in comparison to other diagnostic procedures, is increasing in diagnostic laboratories.