Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Virol. Mar 25, 2024; 13(1): 88164
Published online Mar 25, 2024. doi: 10.5501/wjv.v13.i1.88164
Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus, hepatitis B virus, and human immunodeficiency virus 1
Waleed Abdelgaber Nemr, Radwan K Nashwa
Waleed Abdelgaber Nemr, Department of Radiation Microbiology, National Center for Radiation Research and Technology, Egyptian Atomic Energy Authority, Cairo 11371, Egypt
Radwan K Nashwa, Department of Health Radiation Research, National Center for Radiation Research and Technology, Egyptian Atomic Energy Authority, Cairo 11371, Egypt
Author contributions: Both of the listed authors contributed to the present work equally; both authors read and approved the final manuscript.
Institutional review board statement: The current study has not subjected human or animals to any experiment.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Waleed Abdelgaber Nemr, PhD, Doctor, Department of Radiation Microbiology, National Center for Radiation Research and Technology, Egyptian Atomic Energy Authority, 3th Ahmed Alzomor Street, Cairo 11371, Egypt. wnemr@hotmail.com
Received: September 16, 2023
Peer-review started: September 16, 2023
First decision: November 9, 2023
Revised: December 7, 2023
Accepted: January 10, 2024
Article in press: January 10, 2024
Published online: March 25, 2024
Processing time: 177 Days and 1.1 Hours
Abstract
BACKGROUND

Hepatitis C virus (HCV), hepatitis B virus (HBV), and human immunodeficiency virus 1 (HIV-1) are the most epidemic blood-borne viruses, posing threats to human health and causing economic losses to nations for combating the infection transmission. The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive, but they are more accurate than serological testing.

AIM

To develop a rapid, cost-effective, and accurate diagnostic multiplex polymerase chain reaction (PCR) assay for simultaneous detection of HCV, HBV, and HIV-1.

METHODS

The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electrophoretic molecular weight inside each viral genome. Therefore, this diagnostic method will be appropriate for application in both conventional (combined with electrophoresis) and real-time PCR facilities. Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus. Then, Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay.

RESULTS

The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay. The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis. Compared to related published assays, the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays.

CONCLUSION

This study provides a simple, cost-effective, and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses; this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Keywords: Diagnosis; Blood-borne viruses; Multiplex polymerase chain reaction; High-resolution melting

Core Tip: The current study approaches a cost-effective diagnostic assay to detect the most common blood-borne viruses (hepatitis C virus, hepatitis B virus, and human immunodeficiency virus 1) in a single test of multiplex polymerase chain reaction (PCR). This article includes the procedures in computational biology to achieve the PCR design and the practical examination of the given assay for detecting the targeted viruses with the interpretation of the results.