• Development of a real-time polymerase chain reaction based on SYBR Green dye for rapid detection of common Hepatitis C virus
  • Paria Pirasteh,1 Seyed Reza Mohebbi,2,* Seyed Masoud Hosseini,3 Mohammad Reza Zali,4
    1. Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
    2. Research Center for Gastroenterology and Liver Diseases, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
    3. Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran
    4. Research Center for Gastroenterology and Liver Diseases, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran


  • Introduction: Hepatitis is an inflammation of the liver which mainly happens as a result of viral infections. Hepatitis C virus (HCV) is one of the major public health issues around the world. HCV infection may lead to chronic hepatitis, severe liver damage including liver cirrhosis and hepatocellular carcinoma. Up to now, several various techniques have been developed to detect and monitor HCV infection. However, choosing a rapid and at the same time accurate detection method is a matter of concern. The aim of this study was to develop a SYBR Green dye based real-time polymerase chain reaction (PCR) for HCV detection using an efficient, fast and relatively low-cost method.
  • Methods: Viral RNA genome was extracted from patients’ sera and the cDNA was synthesized. The newly designed specific primers were analyzed by BioEdit and Gene Runner softwares and limiting factors such as the melting temperature probable primer dimers were rechecked and used for amplification of a fragment at the HCV NS5B gene and utilizing real-time PCR technique based on SYBR green chemistry.
  • Results: Positive samples that were tested previously with approved methods (commercial kits and in-house Nested PCR methods), used to assess our developed test. After setting up, similar detection results were observed in compared with previous standard tests. Target region was successfully amplified with appropriate efficiency.
  • Conclusion: Based on the results, we developed a rapid and accurate molecular method for HCV RNA detection and diagnosis. this method can be used as relatively low cost and reliable test to detect and identify of common hepatitis C viruses.
  • Keywords: Primer, real-time PCR, Hepatitis C, Detection