Article
Licensed
Unlicensed
Requires Authentication
Development of a low-cost real-time reverse transcriptase-polymerase chain reaction technique for the detection and quantification of hepatitis C viral load
-
Kiana Shahzamani
Published/Copyright:
March 11, 2010
Abstract
Background: It is necessary to develop a highly specific and sensitive assay to quantify the exact amount of hepatitis C virus (HCV) RNA in blood of patients with hepatitis C. For this reason, a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for quantification of HCV RNA in human plasma was developed.
Methods: A pair of primers as well as hybridization probes were selected. A real-time RT-PCR was set up and optimized. To establish the sensitivity of the assay, a serial dilution of HCV standards and reference sera, including the six major HCV genotypes, was used. The performance of the assay was evaluated with 191 known HCV-RNA positive and 100 negative samples.
Results: The real-time assay had a sensitivity of 50 IU/mL, with a dynamic range of detection between 103 and 106 IU/mL. The coefficients of variation of threshold cycle values in intra- and inter-day-runs were <1.77% and 3.40%, respectively. Measurement of HCV-RNA positive samples yielded reproducible data with 100% specificity.
Conclusions: The high sensitivity, simplicity, reproducibility, wide dynamic range, and low cost of this real-time HCV RNA quantification makes this method especially suitable for monitoring viral load during therapy and tailoring of treatment schedules accordingly.
Clin Chem Lab Med 2010;48:777–84.
Received: 2009-11-21
Accepted: 2010-1-8
Published Online: 2010-03-11
Published in Print: 2010-06-01
©2010 by Walter de Gruyter Berlin New York
You are currently not able to access this content.
You are currently not able to access this content.
Articles in the same Issue
- Editorials
- The “Obamanomics”: a revolution in laboratory diagnostics
- Skeletal muscle biomarkers: not new but still interesting diagnostic tools
- Necessity for high-quality reference materials in the harmonization of laboratory assays
- Reviews
- Rhabdomyolysis: historical background, clinical, diagnostic and therapeutic features
- Biochemical markers of muscular damage
- Minireview
- Interpretation of serum parathyroid hormone concentrations in dialysis patients: what do the KDIGO guidelines change for the clinical laboratory?
- Guidelines and Recommendations
- 2010 Consensus statement on the worldwide standardization of the hemoglobin A1c measurement
- Genetics and Molecular Diagnostics
- Development of a low-cost real-time reverse transcriptase-polymerase chain reaction technique for the detection and quantification of hepatitis C viral load
- Microsatellite polymorphisms in intron 2 of the toll-like receptor 2 gene and their association with susceptibility to pulmonary tuberculosis in Han Chinese
- Evaluation of a panel of circulating DNA, RNA and protein potential markers for pathologies of pregnancy
- General Clinical Chemistry and Laboratory Medicine
- Traceability of values for catalytic activity concentration of enzymes: a Certified Reference Material for aspartate transaminase
- Development and preparation of a new serum protein reference material: feasibility studies and processing
- A WHO Reference Reagent for the Serum Transferrin Receptor (sTfR): international collaborative study to evaluate a recombinant soluble transferrin receptor preparation
- Hyperhomocysteinemia is not associated with reduced bone quality in humans with hip osteoarthritis
- Diagnostic performance of anti-citrullinated peptide antibodies for the diagnosis of rheumatoid arthritis: the relevance of likelihood ratios
- New quantitative electrochemiluminescence method (ECLIA) for interleukin-6 (IL-6) measurement
- Evaluation of cell counting and leukocyte differentiation in cerebrospinal fluid controls using hematology analyzers by the German Society for Clinical Chemistry and Laboratory Medicine
- Reference Values and Biological Variations
- The relationship of coffee and green tea consumption with high-sensitivity C-reactive protein in Japanese men and women
- Cancer Diagnostics
- Potential biomarkers for esophageal carcinoma detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
- Identification of candidate molecular markers predicting chemotherapy resistance in non-small cell lung cancer
- Factors affecting soluble mesothelin related protein levels in an asbestos-exposed population
- Cardiovascular Diseases
- Dynamic changes in N-terminal pro-brain natriuretic peptide in acute coronary syndromes treated with percutaneous coronary intervention: a marker of ischemic burden, reperfusion and outcome
- Performance characteristics of a microfluidic Lab-on-Chip electrophoresis system for high-density lipoprotein (HDL) subfraction separation and measurement
- Letters to the Editor
- Hemolysis correction factor in the measurement of serum neuron-specific enolase
- Serum paraoxonase-1 activity and genetic polymorphisms: common errors in measurement and interpretation of results
- Neuropharmacology of schizophrenia: is there a role for leptin?
- Relationship between serum concentrations of keratin-18 and apoptosis in chronic hepatitis C
Keywords for this article
hepatitis C virus;
hybridization probe;
real-time reverse transcriptase-PCR;
viral load
Articles in the same Issue
- Editorials
- The “Obamanomics”: a revolution in laboratory diagnostics
- Skeletal muscle biomarkers: not new but still interesting diagnostic tools
- Necessity for high-quality reference materials in the harmonization of laboratory assays
- Reviews
- Rhabdomyolysis: historical background, clinical, diagnostic and therapeutic features
- Biochemical markers of muscular damage
- Minireview
- Interpretation of serum parathyroid hormone concentrations in dialysis patients: what do the KDIGO guidelines change for the clinical laboratory?
- Guidelines and Recommendations
- 2010 Consensus statement on the worldwide standardization of the hemoglobin A1c measurement
- Genetics and Molecular Diagnostics
- Development of a low-cost real-time reverse transcriptase-polymerase chain reaction technique for the detection and quantification of hepatitis C viral load
- Microsatellite polymorphisms in intron 2 of the toll-like receptor 2 gene and their association with susceptibility to pulmonary tuberculosis in Han Chinese
- Evaluation of a panel of circulating DNA, RNA and protein potential markers for pathologies of pregnancy
- General Clinical Chemistry and Laboratory Medicine
- Traceability of values for catalytic activity concentration of enzymes: a Certified Reference Material for aspartate transaminase
- Development and preparation of a new serum protein reference material: feasibility studies and processing
- A WHO Reference Reagent for the Serum Transferrin Receptor (sTfR): international collaborative study to evaluate a recombinant soluble transferrin receptor preparation
- Hyperhomocysteinemia is not associated with reduced bone quality in humans with hip osteoarthritis
- Diagnostic performance of anti-citrullinated peptide antibodies for the diagnosis of rheumatoid arthritis: the relevance of likelihood ratios
- New quantitative electrochemiluminescence method (ECLIA) for interleukin-6 (IL-6) measurement
- Evaluation of cell counting and leukocyte differentiation in cerebrospinal fluid controls using hematology analyzers by the German Society for Clinical Chemistry and Laboratory Medicine
- Reference Values and Biological Variations
- The relationship of coffee and green tea consumption with high-sensitivity C-reactive protein in Japanese men and women
- Cancer Diagnostics
- Potential biomarkers for esophageal carcinoma detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
- Identification of candidate molecular markers predicting chemotherapy resistance in non-small cell lung cancer
- Factors affecting soluble mesothelin related protein levels in an asbestos-exposed population
- Cardiovascular Diseases
- Dynamic changes in N-terminal pro-brain natriuretic peptide in acute coronary syndromes treated with percutaneous coronary intervention: a marker of ischemic burden, reperfusion and outcome
- Performance characteristics of a microfluidic Lab-on-Chip electrophoresis system for high-density lipoprotein (HDL) subfraction separation and measurement
- Letters to the Editor
- Hemolysis correction factor in the measurement of serum neuron-specific enolase
- Serum paraoxonase-1 activity and genetic polymorphisms: common errors in measurement and interpretation of results
- Neuropharmacology of schizophrenia: is there a role for leptin?
- Relationship between serum concentrations of keratin-18 and apoptosis in chronic hepatitis C
