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 GB virus C (GBV-C) among Iranian HBV positive patients using PCR-RFLP technique. Background: GBV-C was a member of flaviviridae family and recently was proposed to classify as members of a fourth genus in this family, named Pegivirus and it was suggested that at least one quarter of the world s population has been infected with this virus. GBV-C can be transmitted via 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 semi-nested PCR method. After designing specific primers, the semi-nested PCR was optimized, then sequences of PCR products were analyzed with a 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 a 2% agarose gel 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 is more affordable than the other techniques. Therefore, 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. © 2013 RIGLD.