scholarly journals Fully Automated Detection of Hepatitis C Virus RNA in Serum and Whole-Blood Samples

2002 ◽  
Vol 9 (6) ◽  
pp. 1385-1388 ◽  
Author(s):  
Harald H. Kessler ◽  
Alexandra M. K. Clarici ◽  
Evelyn Stelzl ◽  
Gerhard Mühlbauer ◽  
Elisabeth Daghofer ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Rna Extraction ◽  
Hcv Rna ◽  
Molecular Assay ◽  
Blood Samples ◽  
Molecular Assays ◽  
Roche Molecular Diagnostics ◽  
Whole Blood Samples

ABSTRACT In this study, we established a fully automated molecular assay for qualitative detection of hepatitis C virus (HCV) in serum and whole-blood samples and compared it with conventional molecular assays, including manual HCV RNA extraction protocols. Whole-blood samples were collected from patients with and without chronic HCV infection in EDTA tubes and nucleic acid stabilization tubes (NASTs). Prior to HCV RNA extraction, the HCV Internal Control (IC), derived from the COBAS AMPLICOR HCV test, version 2.0 (Roche Molecular Diagnostics), was added. The new assay was based on an automated extraction protocol on the MagNA Pure LC instrument (Roche Applied Science), followed by automated reverse transcription, amplification, hybridization, and detection on the Cobas Amplicor analyzer (Roche Molecular Diagnostics). The detection limit of the new assay was found to be similar to those of conventional molecular assays. In clinical samples, 100% agreement between the new assay and conventional methods was observed. The introduced amount of IC was detected in 45 of 45 serum samples, 41 of 45 EDTA tube whole-blood samples, and 43 of 45 NAST whole-blood samples. Retesting led to more frequent IC detection. The fully automated molecular assay was found to be suitable for detection of HCV RNA in different kinds of sample materials. It may be recommended for use in the high-throughput routine molecular diagnostic laboratory.

Hepatitis C virus (HCV) RNA quantitation in small-volume whole blood samples

2009 ◽  
Vol 47 (01) ◽  
Author(s):  
T Bruns ◽  
K Steinmetzer ◽  
E Ermantraut ◽  
A Stallmach
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Small Volume ◽  
Hcv Rna ◽  
Blood Samples ◽  
Whole Blood Samples

scholarly journals Effects of Storage and Type of Blood Collection Tubes on Hepatitis C Virus Level in Whole Blood Samples

2001 ◽  
Vol 39 (5) ◽  
pp. 1788-1790 ◽  
Author(s):  
H. H. Kessler ◽  
E. Stelzl ◽  
R. B. Raggam ◽  
J. Haas ◽  
F. Kirchmeir ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Blood Collection ◽  
Blood Samples ◽  
Virus Level ◽  
Whole Blood Samples ◽  
Blood Collection Tubes

scholarly journals Hepatitis C virus RNA quantitation and degradation studies in whole blood samples in vitro

Gut ◽  
2007 ◽  
Vol 56 (2) ◽  
pp. 306-307 ◽  
Author(s):  
J Watson ◽  
S Graves ◽  
J Ferguson ◽  
C D'Este ◽  
R Batey
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Hepatitis C Virus Rna ◽  
Blood Samples ◽  
Virus Rna ◽  
Whole Blood Samples ◽  
Degradation Studies

scholarly journals Hepatitis C Virus RNA Quantitation in Venous and Capillary Small-Volume Whole-Blood Samples

2009 ◽  
Vol 47 (10) ◽  
pp. 3231-3240 ◽  
Author(s):  
T. Bruns ◽  
K. Steinmetzer ◽  
E. Ermantraut ◽  
A. Stallmach
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Small Volume ◽  
Hepatitis C Virus Rna ◽  
Blood Samples ◽  
Virus Rna ◽  
Whole Blood Samples

scholarly journals Hepatitis C virus (HCV) infection and cryoglobulinemia: Analysis of whole blood and plasma HCV-RNA concentrations and correlation with liver histology

Hepatology ◽  
2000 ◽  
Vol 31 (3) ◽  
pp. 737-744 ◽  
Author(s):  
Warren N. Schmidt ◽  
Jack T. Stapleton ◽  
Douglas R. LaBrecque ◽  
Frank A. Mitros ◽  
Patricia A. Kirby ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Hcv Infection ◽  
Liver Histology ◽  
Hcv Rna

scholarly journals Early Prediction of Hepatitis C Virus (HCV) Infection Relapse in Nonresponders to Primary Interferon Therapy by means of HCV RNA Whole-Blood Analysis

2004 ◽  
Vol 39 (12) ◽  
pp. 1754-1760 ◽  
Author(s):  
T. Watkins-Riedel ◽  
P. Ferenci ◽  
P. Steindl-Munda ◽  
M. Gschwantler ◽  
C. Mueller ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Interferon Therapy ◽  
Hcv Infection ◽  
Blood Analysis ◽  
Hcv Rna ◽  
Early Prediction ◽  
Whole Blood Analysis

scholarly journals Automated Specific Capture of Hepatitis C Virus RNA with Probes and Paramagnetic Particle Separation

2000 ◽  
Vol 38 (1) ◽  
pp. 18-21
Author(s):  
Hayato Miyachi ◽  
Atsuko Masukawa ◽  
Toshio Ohshima ◽  
Toru Hirose ◽  
Chaka Impraim ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Hemodialysis Patient ◽  
Rna Extraction ◽  
Magnetic Bead ◽  
Inhibitory Effect ◽  
Hcv Rna ◽  
Specimen Preparation ◽  
Manual Method ◽  
Fluid Separation

ABSTRACT We developed and evaluated a prototype automated specimen preparation instrument for the specific capture of hepatitis C virus (HCV) RNA with probes and magnetic bead-fluid separation. HCV RNA was isolated from serum by lysis of virus particles with a chaotropic agent, followed by hybridization of the RNA with biotinylated probes and capture of the hybridized RNA with streptavidin-coated paramagnetic particles. After washing of the hybrid-particle complexes to remove nonspecifically bound materials, the particles were resuspended in a specimen diluent and were then ready for amplification and detection with a fully automated PCR system (COBAS AMPLICOR; Roche Diagnostic Systems). The analytical sensitivity in the dilution series was 33 copies per ml or greater. Comparison of the test results with those obtained by a manual method based on organic extraction and precipitation of RNA (SepaGene RV-R; Sanko Junyaku Co., Ltd.) showed 93% (49 of 53 samples) sensitivity and 100% (12 of 12 samples) specificity. There was 94% overall agreement between results. When RNA was extracted by the manual method from serum containing 10 3 or 10 5 copies of HCV per ml in the presence of heparin, there was an inhibitory effect on detection of both HCV RNA and the internal control. In contrast, when RNA was extracted from the serum by the automated method, there was no inhibitory effect. This inhibitory effect of heparin on the manual method was also observed for a series of serum specimens from a hemodialysis patient, but the inhibitory effect was eliminated by the automated specimen preparation method. In summary, a fully automated RNA extraction system for PCR detection of HCV RNA by use of specific capture with probes and magnetic bead-fluid separation was shown to have performance similar to that of the conventional manual method. In addition, it successfully eliminated the inhibitory effect of the heparin in the serum and permitted the detection of HCV RNA in serum samples from a hemodialysis patient. The prototype automated RNA extraction system is suitable as a totally automated system, starting with RNA extraction to detection of HCV, if it was combined with the fully automated COBAS AMPLICOR PCR system.

scholarly journals Distribution of Hepatitis C Virus (HCV) RNA in Whole Blood and Blood Cell Fractions: Plasma HCV RNA Analysis Underestimates Circulating Virus Load

1997 ◽  
Vol 176 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Warren N. Schmidt ◽  
Ping Wu ◽  
Jian‐Qiu Han ◽  
Mary Jeanne Perino ◽  
Douglas R. LaBrecque ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Blood Cell ◽  
Whole Blood ◽  
Hcv Rna ◽  
Virus Load ◽  
Rna Analysis ◽  
Cell Fractions

Molecular Detection of Hepatitis C Virus amongst Patients in Five Selected Hospitals in Niger State, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 60-69
Author(s):  
M U Iduh ◽  
F A Kuta ◽  
M E Abalaka ◽  
K O Shitu
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Direct Sequencing ◽  
Rna Extraction ◽  
Public Health Problem ◽  
Developed Countries ◽  
Response To Treatment ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Polymerase Chain

Hepatitis C Virus (HCV) is a major public health problem in developing and developed countries worldwide. It is responsible for liver diseases and hepatocellular carcinoma in chronically-infected patients. This study therefore aimed to identify the strain of HCV among HCV seropositive subjects in Niger State. A total of 44 HCV seropositive blood samples which consisted of 27 males and 17 females were analyzed (after Viral RNA extraction) for the presence of HCV-RNA by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Nine (20.5%) of the samples were positive for HCV RNA. HCV-RNA positive samples were genotyped by direct sequencing at 5’UTR region genomes; sequences were aligned on MEGA 6.0 and confirmed by phylogenetic analysis. HCV genotype 1b was the only one distributed among the participants. The findings are relevant as predictors for using antiviral therapy in this population because the response to treatment varies according to the genotype.

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