A Simple, Sensitive SSCP Method for Detecting HVR1 Quasi-Species of Hepatitis C Virus
Single-strand conformation polymorphism (SSCP) method for detecting of HVR1 quasi-species of HCV is necessary for hepatitis C diagnosis. In this study, we described a rapid and sensitive SSCP method to analyze HCV quasi-species using common equipment. After RT-PCR reaction, denaturing was conducted with heating and Chilling. By varying the electrophoresis temperature, voltage and total acrylamide concentration，the extent of cross-linking in the gels (1-2.5%) and the concentration in glycerol ( 0-10％), SSCP method was established. With the optimized procedure, variants representing 2-3% of the whole quasi-species could be detected. By isolating visible bands from electrophoresis gel followed by sequencing, the genetic variability of HCV can be concluded. This method has high accuracy and reproducibility, simplicity and low cost, which could be a useful tool in laboratory practice.
T. L. Jia et al., "A Simple, Sensitive SSCP Method for Detecting HVR1 Quasi-Species of Hepatitis C Virus", Advanced Materials Research, Vols. 343-344, pp. 568-575, 2012