Prevalence of GBV-C among Iranian HBV positive patients using PCR-RFLP technique

Aim: The aim of this study was to investigate the prevalence of GBV-C among Iranian HBV positive patients using
PCR-RFLP technique.
Background: GBV-C was a member of flaviviridae family and recently propose to classify as members of a fourth
genus in this family, named Pegivirus and suggest that at least one quarter of the world's population has been infected
with this virus. GBV-C can be transmitted via the blood-borne route, although vertical and sexual transmission is very
well documented.
Patients and methods: 100 serum samples were collected from HBsAg positive patients in 2011-2012. RNA was
extracted with Qiagene mini kit. cDNA was synthesized by Reverse Transcriptase method and amplified by Seminested
PCR method .After designing specific primers, the semi nested PCR was optimized, then sequences of PCR
products were analyzed with software such as neb cutter, and sites of restriction enzymes were determined and suitable
enzymes were selected for RFLP (Restriction Fragment Length Polymorphism).
Results: PCR products were analyzed in 2% agarose gels containing ethidium bromide and were visualized with
ultraviolet (UV) light. A 230 bp band was observed in comparison with 100 kb ladder; this band indicates our target
gene of GBV-C genome have been isolate from serum samples.
Conclusion: It seems that Co-infection of GBV-C and HBV are common and This method had acceptable sensitivity for
detecting GBV-C and determining its genotype, and more affordable than the other techniques; so the results of this
study showed the prevalence of GBV-C were 12 serums of 100 serums HBsAg positive in goal population and one
sample from 12 GBV-C positive serums was genotype 3 and the others were genotype 2