Author/Authors :
Shahzamani, Kiana Digestive Disease Research Center - Tehran University of Medical Sciences, Tehran , Sabahi, Farzaneh Department of Virology - Tarbiat Modarres University, Tehran , Merat, Shahin Digestive Disease Research Center - Tehran University of Medical Sciences, Tehran , Sadeghizadeh, Majid Department of Genetics - Tarbiat Modarres University, Tehran , Esmail Lashkarian, Hamed Department of Genetics - Lorestan University of Medical Sciences, Khorramabad , Rezvan, Houri Molecular Laboratory - Day General Hospital, Tehran , Mirab Samiee, Siamak Food and Drug Control Research Laboratories, Tehran , Karimi Arzanani, Mohsen Department of Biotechnology - Pasteur Institute of Iran, Tehran , Jabbari, Hossain Digestive Disease Research Center - Tehran University of Medical Sciences, Tehran , Malekzadeh, Reza Digestive Disease Research Center - Tehran University of Medical Sciences, Tehran
Abstract :
Background: We intend to design and validate a low-cost assay for the detection of hepatitis C virus (HCV) RNA using rapid-cycle RTPCR.
The procedure is performed in a closed system with little risk of contamination allowing PCR and product identification to be performed
within one or two hours.
Methods: A SYBR Green-based real-time RT-PCR for rapid detection of HCV. Amplicon synthesis was monitored continuously by SYBR
Green I, which binds to double stranded DNA during PCR. The PCR products were identied by melting curve analysis. Standard sera with
known concentrations of HCV RNA and 150 clinical samples were used to validate our assay.
Results: The minimum detection level of our assay was less than 50 IU/mL. The results on 100 plasma samples were comparable with
commercial assays.
Conclusions: This method is useful for rapid qualitative detection of HCV infection and particularly suitable for routine diagnostic applications.
Keywords :
hepatitis C virus , rapid detection , real-time RT-PCR , SYBR Green I
