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
ARTICLE HIGHLIGHTS
Research background

The most epidemic blood-borne viruses are hepatitis C virus (HCV), hepatitis B virus (HBV), and human immunodeficiency virus 1 (HIV-1). They cause mortality for millions of people worldwide. Although serology diagnostic methods are less accurate than nucleic acid amplification testing (NAT) for detecting blood-borne viruses in blood samples, they are commonly used to save money and time.

Research motivation

The innovation of rapid and cost-effective NAT assays will allow us to substitute the mistakable serologoical assays, and this can be achieved through a multiplex polymerase chain reaction (PCR) assay for the simultaneous detection of blood-borne viruses in a single test.

Research objectives

The present study focused on developing a new multiplex PCR assay for simultaneous detection of HCV, HBV, and HIV-1 in a single tube.

Research methods

The in silico design of the PCR assay targets conserved sequences in each viral genome among all variants. This was evaluated by multiple sequence alignment and finding the lowest entropy regions. The selected primers were evaluated also to avoid the possibility of forming stable dimers or hairpins. All primers were harmonized in the melting temperature to anneal at the same temperature during PCR. A practical experiment was conducted to prove the feasibility of the present assay.

Research results

The in silico evaluations proved the worthiness of the selected primers to match with many variants of each virus with negligible ability to form dimers, this ensures the efficiency of the proposed PCR assay. Consequently, the sensitivity of this assay showed the ability to detect HCV at a limit of detection (LOD) of 61.5 IU/mL. Furthermore, the LOD value is 7 IU/mL for HBV and 98.3 IU/mL for HIV-1.

Research conclusions

The proposed cost-effective PCR assay of the current study achieved a competitive sensitivity with the analogous multiplex PCR assays.

Research perspectives

The findings of the current study encourage further developmental studies to apply this assay in an automated system for large-scale virology screening of blood samples.