Type-Specific Detection of Human Papillomaviruses in a Routine Laboratory Setting – Improved Sensitivity and Specificity of PCR and Sequence Analysis Compared to Direct Hybridisation
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Siegfried Kösel
Abstract
Human papillomaviruses (HPV) are known to cause cervical dysplasia and cervical carcinoma. We used a 3-step PCR protocol that allows rapid type-specific HPV testing in a routine laboratory setting: HPV-16-positive samples were determined using a specific Light-Cycler PCR; HPV-16-negative samples were amplified by nested PCR and typed by sequence analysis. During a period of 7 months, 1275 PCR-based HPV tests were performed. Of the 1275 samples, 829 samples tested negative for HPV and 446 tested positive, including 124 positives found in the initial HPV-16-specific Light-Cycler assay. Sequence analysis of 132 samples detected 18 HPV types that are not included in the widely used Hybrid Capture II assay. For comparison, the first 100 cervical specimens were tested in parallel using PCR and direct hybridisation (Hybrid Capture II assay). PCR detected HPV DNA in 23 samples that tested negative in the Hybrid Capture assay. Four out of 37 samples that tested positive for HPV in the Hybrid Capture test may be false positives, because sequence analysis detected HPV types not included in the probe mixtures. As rare and novel HPV types may also confer an oncogenic risk, highly sensitive and specific PCR assays will help in understanding cervical HPV infection and cervical cancer of unknown causes.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Editorial
- Biomedical Applications of Capillary Electrophoresis
- Capillary Electrophoresis – A High Performance Analytical Separation Technique
- The Analysis of Carbohydrate-Deficient Transferrin, Marker of Chronic Alcoholism, Using Capillary Electrophoresis
- Capillary Electrophoresis of Hemoglobin
- Capillary Electrophoresis for the Determination of Organic Acidurias in Body Fluids: A Review
- Separation of Serum Proteins by Automated Capillary Zone Electrophoresis
- Is Capillary Electrophoresis a Method of Choice for Systematic Toxicological Analysis?
- Capillary Zone Electrophoresis of Serum Proteins: Effects of Changed Analytical Conditions
- Type-Specific Detection of Human Papillomaviruses in a Routine Laboratory Setting – Improved Sensitivity and Specificity of PCR and Sequence Analysis Compared to Direct Hybridisation
- Identification of Lipoproteins of Intestinal Origin in Human Atherosclerotic Plaque
- Molecular Cloning, Sequencing and Functional Expression of Porcine Thyrotropin (TSH) Receptor cDNA
- Are Results of Fibrinogen Measurements Transferable?
- Comparison of Several Point-of-Care Testing (POCT) Glucometers with an Established Laboratory Procedure for the Diagnosis of Type 2 Diabetes Using the Discordance Rate. A New Statistical Approach
- Association between Increased Serum Cholesterol and Signs of Depressive Mood
- Report from the Second European Symposium on Clinical Laboratory and in vitro Diagnostic Industry. Physiological Reference Values: A Shared Business? Barcelona, 67 February 2003
- Meetings and Awards
Articles in the same Issue
- Editorial
- Biomedical Applications of Capillary Electrophoresis
- Capillary Electrophoresis – A High Performance Analytical Separation Technique
- The Analysis of Carbohydrate-Deficient Transferrin, Marker of Chronic Alcoholism, Using Capillary Electrophoresis
- Capillary Electrophoresis of Hemoglobin
- Capillary Electrophoresis for the Determination of Organic Acidurias in Body Fluids: A Review
- Separation of Serum Proteins by Automated Capillary Zone Electrophoresis
- Is Capillary Electrophoresis a Method of Choice for Systematic Toxicological Analysis?
- Capillary Zone Electrophoresis of Serum Proteins: Effects of Changed Analytical Conditions
- Type-Specific Detection of Human Papillomaviruses in a Routine Laboratory Setting – Improved Sensitivity and Specificity of PCR and Sequence Analysis Compared to Direct Hybridisation
- Identification of Lipoproteins of Intestinal Origin in Human Atherosclerotic Plaque
- Molecular Cloning, Sequencing and Functional Expression of Porcine Thyrotropin (TSH) Receptor cDNA
- Are Results of Fibrinogen Measurements Transferable?
- Comparison of Several Point-of-Care Testing (POCT) Glucometers with an Established Laboratory Procedure for the Diagnosis of Type 2 Diabetes Using the Discordance Rate. A New Statistical Approach
- Association between Increased Serum Cholesterol and Signs of Depressive Mood
- Report from the Second European Symposium on Clinical Laboratory and in vitro Diagnostic Industry. Physiological Reference Values: A Shared Business? Barcelona, 67 February 2003
- Meetings and Awards
