RAPID AND SENSITIVE METHOD FOR DETECTION OF HEPATITIS-C VIRUS-RNA BY USING SILICA PARTICLES

We describe a rapid, sensitive, and economic method for detection of h epatitis C virus (HCV) RNA. This method uses silica particles for puri fication of nucleic acid and then a modified reverse transcription-PCR that minimizes the risk of contamination and reduces the amount of re agents used. We found purification by silica particles to be at least as sensitive and in certain circumstances more sensitive than that by traditional phenol-chloroform extraction. This improved sensitivity ma y be due to more efficient recovery of HCV RNA by silica particles. HC V RNA appears to bind to silica particles in a saturable fashion, and the addition of extraneous nucleic acids (salmon sperm DNA or tRNA) de creases the binding in a dose-related fashion. The reverse transcripti on-PCR is performed by using a modified single tube method which furth er simplifies and reduces the cost of this assay. Finally, this method may be applied to clinical specimens such as liver tissue.