Development of a SYBR green real time multiplex RT-PCR technique for simultaneous detection of HCV and GBV-C Co-infection in plasma samples

Background:Accumulative research is in progress to clarify clinical aspects of GBV-C. The possibility of interaction between HCV and GBV-C as well as its consequence on development of liver diseases is the most important clinical aspect which encourages researchers to develop a rapid and cost effective technique for simultaneous detection of both viruses. Methods: In this study, a SYBR Green real time multiplex RT-PCR technique as a new economical and sensitive method was designed and validated for simultaneous detection of HCV/GBV-C in HCV positive plasma samples. SYBR green real time RT-PCR technique optimization was performed separately for each virus. Multiplex PCR was established next. Standard sera with known concentrations of HCV RNA and dual HCV/GBV-C positive control samples along with negative control samples were used to validate the assay. Results and Conclusions:  Fifty six non cirrhotic HCV positive plasma samples [۲۹ of genotype ۳a and ۲۷ of genotype ۱a] were collected from patients before receiving treatment. ۲۰.۶% of genotype ۳a and ۱۸.۷% of genotype ۱a showed HCV/GBV-C co-infection. As a result, ۱۹.۶% of ۵۶ samples had HCV/GBV-C co-infection that was compatible with other results from all over the world. SYBR Green real time multiplex RT-PCR technique can be used to detect HCV/GBV-C co-infection in plasma samples. Furthermore, with application of this method more time and cost could be saved in clinical-research settings.

کلیدواژه ها:

HCV ، GBV-C ، SYBR Green real time ، Rapid detection ، multiplex RT- PCR