Detectable Serum HCV-RNA Research Articles

Single-step reverse transcription-polymerase chain reaction for hepatitis C virus RNA with DNA enzyme immunoassay hybridization

A single reverse transcription-polymerase chain reaction (SRT-PCR) for HCV RNA detection, confirmed by hybridization of amplified products with biotinylated probes using DNA enzyme immunoassay (DEIA), was compared to nested-PCR (N-PCR) on 20 sera from patients with chronic (n = 18) or acute (n = 2) hepatitis. Results obtained by SRT-PCR confirmed by DEIA and by N-PCR identical. All but one of the patients with chronic hepatitis and positive HCV serology as well as the patients with acute hepatitis had detectable HCV RNA in serum; all patients with chronic hepatitis and indeterminate HCV serology and all controls (n = 5) were negative by the two PCR methods. Both SRT-PCR and N-PCR remained positive after 7 x 10(-2) and 5 x 10(-4) dilutions of two HCV RNA-positive sera. The threshold of detection for SRT-PCR was 15 RNA copies per assay, as assessed by testing serial dilutions of an in vitro synthesized 5'-NCR HCV RNA transcript. SRT-PCR confirmed by DEIA for HCV RNA appears to be as sensitive and specific as N-PCR. Furthermore, it is easier to perform, with less of contamination, is less time-consuming, requires fewer enzymes, and it permits semi-quantification of PCR products.

Detection and quantitation of serum HCV-RNA by branched DNA amplification in anti-HCV positive blood donors

The detection of serum HCV-RNA needs to be standardized. The aim of this study was to assess the effectiveness of the branched DNA amplification method in detecting and quantitating serum HCV-RNA in 54 blood donors, 33 with and 21 without increased serum alanine aminotransferase levels and with detectable serum HCV-RNA by polymerase chain reaction. HCV-RNA was detected by branched DNA signal amplification in 42/54 (77%) of the blood donors. Positivity rates were not different among the 21 blood donors with normal and the 33 blood donors with increased serum alanine aminotransferase levels (86% and 76%, respectively). Median serum HCV-RNA levels were not different among donors with or without increased serum alanine aminotransferase levels (28.6 x 10(5) Eq/ml and 14.7 x 10(5) Eq/ml, respectively). There was no significant correlation between serum alanine aminotransferase levels and serum HCV-RNA levels. These findings show that branched DNA signal amplification identifies most of the donors with true hepatitis C virus viremia and that the level of hepatitis C virus replication is not correlated to serum alanine aminotransferase levels.

直接ＲＴ‐ＰＣＲ法による血清からのＨＣＶ ＲＮＡ迅速検出

直接ＲＴ‐ＰＣＲ法による血清からのＨＣＶ ＲＮＡ迅速検出

Detection of serum HCV-RNA by PCR in anti-HCV positive patients

Detection of serum HCV-RNA by PCR in anti-HCV positive patients

Detection of serum HCV-RNA in anti-HCV positive subjects with normal ALT level correlates with liver HCV-RNA and liver histology

Detection of serum HCV-RNA in anti-HCV positive subjects with normal ALT level correlates with liver HCV-RNA and liver histology

Markers of hepatitis C virus infection in Sardinian blood donors: relationship with alanine aminotransferase levels.

Serum samples from 1,765 consecutive Sardinian blood donors, negative for hepatitis B surface antigen (HBsAg) and for antibodies to human immunodeficiency virus (HIV) (anti-HIV), were evaluated for the presence of antibodies to hepatitis C virus (anti-HCV) by second-generation ELISA. Anti-HCV was detected in 25 (1.45%) of the 1,765 donors examined. Anti-HCV was found in 15 of the 1,690 (0.9%) donors with normal alanine aminotransferase (ALT) and in 10 of the 75 (13%) donors with elevated ALT (P < 0.0001). Of the 15 anti-HCV-positive donors with normal ALT, only five (33%) were confirmed to be positive by second-generation RIBA, six (40%) were indeterminate, while four (27%) were RIBA negative. HCV RNA, as detected by polymerase chain reaction (PCR) using a set of primers from the 5'-noncoding region, was found in six of the 15 (40%) donors with normal ALT, including five RIBA positive and one indeterminant. Of the 10 anti-HCV-positive donors with elevated ALT, all were RIBA positive and eight (80%) had detectable HCV RNA. Thus, among ELISA-reactive donors, those with elevated ALT had a significantly higher probability of being positive for second-generation RIBA and HCV RNA compared to those with normal ALT levels (P = 0.028). None of the 65 donors with elevated ALT but negative for anti-HCV by ELISA had detectable serum HCV RNA, as compared to eight of 10 anti-HCV ELISA-positive donors (P < 0.0001). However, although negative for HBsAg, 12 of the 65 (18%) had serum HBV DNA by PCR.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of hepatitis C virus RNA in serum and peripheral blood mononuclear cells

Hepatitis C virus (HCV) RNA is the most reliable marker of HCV infection. However, because of HCV genome heterogeneity and the procedures used to isolate the viral genome and to perform the amplification steps, results do not accurately reflect the actual rate of HCV infection. In this study, several isolation procedures and sets of primers to perform PCR were compared for the detection of HCV-RNA in serum and peripheral blood mononuclear cells (PBMC) of patients with HCV infection. It was found that the extraction with guanidinium isothiocyanate prior to cDNA synthesis followed by PCR amplification of the 5'-UTR region gave the best results in serum and PBMC. The presence of the viral genome in PBMC is a frequent event during HCV infection and this has important consequences for the understanding of the viral pathogenesis.

Detection of HCV-RNA in serum of patients with chronic manb hepatitis using synthetic oligonucleot1des

Detection of HCV-RNA in serum of patients with chronic manb hepatitis using synthetic oligonucleot1des