Qualitative detection of GB Virus C and Hepatitis C Virus co-infection in cirrhotic patients using a SYBR green multiplex RT-PCR technique.

GB Virus C (GBV-C) and Hepatitis C Virus (HCV) belong to the Flaviviridae family of viruses and GBV-C is the closest virus to HCV genetically. Possibility of interaction between HCV and GBV-C and its association with other liver diseases are the most important clinical aspects which encourage researchers to develop a technique for detection of these viruses simultaneously. In this study a SYBR Green multiplex real time RT-PCR technique as a new economical and sensitive method was optimized for simultaneous detection of HCV/ GBV-C in cirrhotic patients. After designing two pairs of specific primers for HCV and GBV-C, SYBR Green Real time RT-PCR technique optimization was performed separately for each virus. Then, multiplex PCR was developed. Finally the optimized technique was performed on positive and negative plasma samples. Eighty nine cirrhotic HCV positive plasma samples (29 of genotype 3a and 27 of genotype 1a) were collected from patients before receiving treatment. 14% of genotype 3a and 17.1% of genotype 1a showed HCV/ GBV-C co-infection. As a result, 13.48% of 89 samples had HCV/ GBV-C co-infection that was compatible with other results from all over the world. Data showed no apparent influence of HGV co-infection on the either clinical or virological aspect of HCV infection. Furthermore, with application of multiplex Real time RT-PCR technique, more time and cost could be saved in clinical-research settings.