Evaluation of Multiplex Nested Polymerase Chain Reaction for Routine Hepatitis C Virus Genotyping in Egyptian Patients

Mohamed Shemis; Dina El-Abd; Dalia Ramadan; Mohamed El-Sayed; Bassem Shenoda Guirgis; Mohamed Saber; Hassan Said Azzazy

doi:10.5812/hepatmon.830

Hepatitis Monthly

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https://doi.org/10.5812/hepatmon.830

Evaluation of Multiplex Nested Polymerase Chain Reaction for Routine Hepatitis C Virus Genotyping in Egyptian Patients

Author(s):

Mohamed Abbas Shemis¹,

Dina Mohamed El-Abd²,

Dalia Ibrahim Ramadan^3,*,

Mohamed Ibrahim El-Sayed²,

Bassem Shenoda Guirgis²,

Mohamed Ali Saber⁴,

Hassan Mohamed El- Said Azzazy²

1Department of Biochemistry, Theodor Bilharz Research Institute, Egypt

2Department of Clinical and Chemical Pathology, Faculty of Medicine, Cairo University, Egypt

3Department of Clinical and Chemical Pathology, Faculty of Medicine, Cairo University, [email protected], Egypt

4Department of Biochemistry, Theodor Bilharz Research Institute, Egypt

Hepatitis Monthly:Vol. 12, issue 4; 265-270

Published online:Apr 30, 2012

Article type:Research Article

Received:Nov 03, 2011

Accepted:Jan 25, 2012

How to Cite:Shemis M, El-Abd D, Ramadan D, El-Sayed M, Shenoda Guirgis B, et al. Evaluation of Multiplex Nested Polymerase Chain Reaction for Routine Hepatitis C Virus Genotyping in Egyptian Patients. Hepat Mon. 2012;12(4):. doi: https://doi.org/10.5812/hepatmon.830

Abstract

Background: At least six HCV (hepatitis C virus) genotypes are unequally distributed worldwide. HCV genotyping guides the selection of treatment regimens and provides important epidemiological markers that enable the outbreak source to be traced and the spread of disease to be controlled. In Egypt, there is an increasing need for cost-effective, fast, and easily performable HCV genotyping assays.Recently, a multiplex PCR assay was developed to determine HCV genotypes. It employs genotype-specific primers, based on sequences of the entire core region and part of the 5’UTR of the genome.
Objectives: In this study, we compared a simple, new, modified multiplex PCR system for HCV genotyping with a commercially available line probe assay (INNO-LiPA) that is based on reverse hybridization.
Patients and Methods: Serum samples from chronic HCV Egyptian patients (n = 73) were genotyped using the modified multiplex PCR assay, and genotypes were verified using the INNO-LiPA HCV II assay.
Results: The modified multiplex PCR method was able to type HCV-4 in 65 of 70 typeable samples (92.86%) and had 100% concordance with the INNO-LiPA assay.
Conclusions: Genotype 4 was the most prevalent genotype in our study. Based on our results, the modified multiplex nested PCR assay is a sensitive and inexpensive alternative for HCV genotyping and can be used in routine diagnostic laboratories. INNO-LiPA may be useful as a second-line assay for genotyping samples that are indeterminate by multiplex PCR. This approach will effect better treatment optimization and a reduction of the spread of HCV.

Implication for health policy/practice/research/medical education:
Egypt has the highest prevalence of worldwide HCV (15%) and the highest prevalence of HCV-4, which is responsible for almost 90% of infections. There is increasing evidence that patients with different HCV genotypes have different clinical profiles, severity of liver disease and response to current combination therapy. HCV genotyping helps clinicians in their practice as it guides the selection of treatment regimens, and provides important epidemiological markers that enable outbreak source tracing and control of the spread of disease.

Please cite this paper as:
Shemis MA, El-Abd DM, Ramadan DI, El-Sayed MI, Guirgis BS, Saber MA, et al. Evaluation of Multiplex Nested Polymerase Chain Reaction for Routine Hepatitis C Virus Genotyping in Egyptian Patients. Hepat Mon. 2012;12(4): 265-270. DOI: 10.5812/hepatmon.830

Keywords

Hepatitis C

Multiplex Polymerase Chain Reaction

Branched DNA Signal Amplification

Assay

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Author(s):

Mohamed Shemis:[PubMed][Scholar]
Dina El-Abd:[PubMed][Scholar]
Dalia Ramadan:[PubMed][Scholar]
Mohamed El-Sayed:[PubMed][Scholar]
Bassem Shenoda Guirgis:[PubMed][Scholar]
Mohamed Saber:[PubMed][Scholar]
Hassan Said Azzazy:[PubMed][Scholar]

Official Journal of Research Center for Gastroenterology and Liver Diseases

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Evaluation of Multiplex Nested Polymerase Chain Reaction for Routine Hepatitis C Virus Genotyping in Egyptian Patients

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