Evaluation of multiplex nested polymerase chain reaction for routine hepatitis C virus genotyping in egyptian patients.
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.


OBJECTIVE
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.


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.
Department of Biochemistry, Theodor Bilharz Research Institute, Giza, Egypt.
Mohamed Abbas
MA
Shemis
Dina Mohamed
DM
El-Abd
Dalia Ibrahim
DI
Ramadan
Mohamed Ibrahim
MI
El-Sayed
Bassem Shenoda
BS
Guirgis
Mohamed Ali
MA
Saber
Hassan Mohamed El-Said
HM
Azzazy