Analysis of hepatitis C virus RNA prevalence and surrogate markers of infection among seropositive voluntary blood donors

Abstract

A novel reverse transcription polymerase chain reaction assay has been developed that uses drop-in-drop-out primers for the heminested amplification of hepatitis C virus complementary DNA. This assay has been used for analysis of the prevalence of hepatitis C virus RNA in a set of 53 plasma specimens from blood donations that were repeatedly reactive for hepatitis C virus antibodies with the first-generation enzyme immunoassay. Of 21 specimens that were also reactive for hepatitis C virus antibodies by a four-antigen recombinant immunoblot assay (recombinant immunoblot assay 2), 20 (95%) contained detectable levels of hepatitis C virus RNA. Cryoprecipitate in three specimens reactive in the recombinant immunoblot assay led to an apparent failure of detecting hepatitis C virus RNA, but repeat tests of redissolved cryoprecipitate subsequently revealed hepatitis C virus RNA. Hepatitis C virus RNA was also detected in plasma from 5 of 29 donors nonreactive by recombinant immunoblot assay. However, evidence of viremia in these donors could not be confirmed on follow-up specimens collected more than 1 yr later. Our results demonstrate that the presence of recombinant immunoblot assay reactivity nearly always indicates hepatitis C viremia and suggest that viremia may be transient or fluctuating among some individuals who are nonreactive for hepatitis C virus antibodies by the recombinant immunoblot assay.

Assessment of Viral Genotype Impact to the Cost-Effectiveness and Overall Costs of Care for Peg-Interferon-2α + Ribavirine Treated Chronic Hepatitis C Patients

“…Our observations regarding stage of disease and comorbidities impact to patterns of health services utilization and overall costs were mostly in line with the findings of Wong et al (13). Authors observed statistically significant differences between groups for few clinical outcomes too, such as AST enzyme and bilirubin, both widely accepted surrogate markers of chronic hepatitis activity (28). Particularly evident difference was noticed for viral RNA decrease level in favor of the genotype group II, contingent with the findings of Singal et al (29).…”

Detection of Hepatitis C Virus RNA: Application to Diagnostics and Research

“…Two distinct procedures for detection of PCR products with 32 P-end-labelled oligonucleotide(s) have been published: agarose gel electrophoresis, followed by Southern blotting and filter hybridization with end-labelled probe(s) [15,25,27,31], and liquid hybridization with labelled oligonucleotide(s) and subsequent separation of the hybridization hybrids from the nonhybridized probe [46, 48,49]. Both procedures have the same sensitivity [46,48].…”

“…To reach a higher sensitivity of HCV-RNA detection, the virus has been concentrated from larger volumes of plasma or serum (up to 8 ml) by ultracentrifugation or polyethylene glycol (PEG) precipitation in some of the published protocols for cDNA-PCR [27,30,31]. Comparison of the efficacy of these procedures revealed that ultracentrifugation improves the sensitivity of the cDNA-PCR assay, resulting in a 2 log higher detection of HCV-RNA [31]. However, ultracentrifugation also induces a significant risk of crosscontamination.…”

“…Therefore, amplification of HCV-RNA by cDNA-PCR is in most of the protocols developed in-house a two-step procedure requiring separate enzymes and buffer conditions for the cDNA reaction and PCR [25,27,31,34,35]. Precipitation of the cDNA reaction and subsequent resuspension of the cDNA product in PCR buffer highly increases the risk of contamination [15,16,25,27,31,34]. To avoid this risky step, a number of procedures have been developed that combine cDNA reaction and PCR amplification.…”

Safety considerations in IGIV utilization

“…In 1992, the FDA regulations provided for mandatory testing for hepatitis C antibody and to eliminate plasma with a positive test [8]. There was concern whether the presence of this antibody could act as a neutralizing antibody for the hepatitis C virus [9,10]. In February 1994, both Gammagard and PolygamR were voluntarily withdrawn from the market due to suspected cases of hepatitis C transmission.…”