Detection of hepatitis C viral RNA: comparison of Amplicor single PCR and nested set PCR processes with HCV serology for hemodialyzed patients.
Multicentre quality control studies demonstrated that optimization and standardization of HCV RNA reverse transcription polymerase chain reaction (RT-PCR) amplification techniques were possible: thus, a nested HCV RNA RT-PCR assay was described as a reliable tool for the detection of hepatitis C viral RNA. Besides this, another procedure, the Amplicor HCV RNA qualitative test, a standardized RT-PCR assay, became available. In order to assess the relationship between seropositivity and potential infectivity and to compare both RT-PCR assays, all patients undergoing maintenance hemodialysis were evaluated at hospital de Meaux (Meaux, France) during one year. We conclude that both assays are equally sensitive and that acid guanidinium thiocyanate based methods, which are used to prepare RNA prior to PCR, are more efficient than the usual phenol extraction protocols. Care should be taken with sera from hemodialyzed patients as the presence of inhibitors of the PCR has been demonstrated during the course of HCV RNA testing.
Bogard
M
M
Unité de Biologie Moléculaire, CH de Meaux, France.
Louvet
M
M
Kamalodine
T
T
Barbanel
C
C
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005069:Evaluation Studies as Topic; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D018937:Hepatitis C Antibodies / Q000097:blood; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D006435:Renal Dialysis; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D017931:DNA Primers; D018937:Hepatitis C Antibodies; D012367:RNA, Viral