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 hepatitis C virus (HCV) RNA. This method uses silica particles for purification of nucleic acid and then a modified reverse transcription-PCR that minimizes the risk of contamination and reduces the amount of reagents 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 may be due to more efficient recovery of HCV RNA by silica particles. HCV RNA appears to bind to silica particles in a saturable fashion, and the addition of extraneous nucleic acids (salmon sperm DNA or tRNA) decreases the binding in a dose-related fashion. The reverse transcription-PCR is performed by using a modified single tube method which further simplifies and reduces the cost of this assay. Finally, this method may be applied to clinical specimens such as liver tissue.
Cheung
R C
RC
Department of Medicine, Palo Alto VA Medical Center, CA 94304.
Matsui
S M
SM
Greenberg
H B
HB
D001483:Base Sequence; D016174:Hepacivirus / Q000235:genetics*; D006508:Hepatitis Antibodies / Q000032:analysis; D018937:Hepatitis C Antibodies; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide
D006508:Hepatitis Antibodies; D018937:Hepatitis C Antibodies; D012367:RNA, Viral; D012822:Silicon Dioxide