In situ reverse transcriptase-polymerase chain reaction: an innovative tool for hepatitis C virus RNA detection and localisation, and for quantification of infected cells.
In situ reverse transcriptase-polymerase chain reaction is a novel and exciting technique which couples the extremely high sensitivity of DNA amplification with the advantages of in situ hybridisation, allowing the preservation of cell morphology and the localisation of the positive signal within intact cells. In this brief report, we analyse and discuss the application of such a technique to the study of hepatitis C virus (HCV) infection, the most common cause of chronic liver disease worldwide. Moreover, given the lymphotropism of this virus, we describe here our own approach to detect and localise HCV RNA within intact peripheral blood mononuclear cells and to quantify the relative number of infected cells.
Muratori
L
L
Department of Internal Medicine, Cardioangiology, Hepatology University of Bologna, Policlinico S. Orsola, Italy. muratori@med.unibo.it
D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization; D012367:RNA, Viral / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase