Quantitative PCR
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Abstract
The classic molecular biology methods like Northern or Southern blot analyse non-amplified DNA or RNA, but need large amounts of nucleic acids, in many instances from tissues or cells that are heterogeneous. In contrast, polymerase chain reaction (PCR)-based techniques allow us to obtain genetic information through the specific amplification of nucleic acid sequences starting with a very low number of target copies. These reactions are characterized by a logarithmic amplification of the target sequences i.e. increase of PCR copies followed by a plateau phase showing a rapid decrease to zero of copy number increment per cycle. Accordingly, the amount of specific DNA product at the end of the PCR run bears no correlation to the number of target copies present in the original specimen. However, many applications in medicine or research require quantification of the number of specific targets in the specimen. This has generated a rapidly increasing need for the development of quantitative PCR techniques. Prominent examples are the determination of viral load in blood specimens for the diagnosis of HIV or HCV infections, the determination of changes in gene dosage through amplification or deletion e.g. of MDR-1, erb-B2, c-myc or the loss of heterozygosity in general. Finally, the analysis of gene expression on the mRNA level does require quantitative approaches to reverse transcriptase PCR, e.g. for studies in morphogenesis or the profiling of cancer cells. Recent advances in technology allow detection of the increment per cycle of a specifically generated PCR product in “real-time mode”. Together with the new powerful methods to dissect heterogeneous tissues or fractionate bodily fluids, this now sets the stage for a detailed analysis not only of the genes and genetic changes within a single cell, but also of the use such cell makes of its genes e.g. in pharmacogenomics. Examples of recent developments of the technology and their applications will be given.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- The Basis of the Medicine of Tomorrow "Validating and Using Pharmacogenomics" Joint IFCC-Roche Diagnostics Conference, Kyoto, Japan, 1619 April 2000
- Diagnostics and the Future of Medicine
- Operomics: Molecular Analysis of Tissues from DNA to RNA to Protein
- Idiosyncratic Reactions to Drugs: Can Medicine Response Profiles Provide a Dynamic Drug Surveillance System?
- Hunting for Disease Genes in Multi-Functional Diseases
- Familial Studies on the Genetics of Cardiovascular Diseases: the Stanislas Cohort
- Quantitative PCR
- Gene Amplification as Means for Determining Therapeutic Strategies in Human Cancers
- Apolipoprotein E Polymorphisms and Concentration in Chronic Diseases and Drug Responses
- Angiotensin I-Converting Enzyme Gene Polymorphism and Drug Response
- Drug-Metabolizing Enzymes, Polymorphisms and Interindividual Response to Environmental Toxicants
- Database Analysis and Gene Discovery in Pharmacogenetics
- How to Manage Individualized Drug Therapy: Application of Pharmacogenetic Knowledge of Drug Metabolism and Transport
- P-Glycoprotein and Bioavailability-Implication of Polymorphism
- Cancer Therapy and Polymorphisms of Cytochromes P450
- Polymorphisms in UDP Glucuronosyltransferase Genes: Functional Consequences and Clinical Relevance
- The Human Multidrug Resistance-Associated Protein (MRP) Gene Family: From Biological Function to Drug Molecular Design
- Ethnic Differences in Drug Metabolism
- Hypervariable Region 1 of Hepatitis C Virus Genome and Response to Interferon Therapy
- A Functional Genomic Study of the Effects of Antipsychotic Agent Chlorpromazine in PC12 Cells
- Influence of Glutathione S-Transferase M1 and T1 Genotypes on Larynx Cancer Risk among Korean Smokers
- CYP2D6 Genotyping in Patients on Psychoactive Drug Therapy
- Genotyping of CYP2D6 in Parkinsons's Disease
- Rapid Analysis of CGG Repeat Length in the FMR1 Gene
- Multiplex In-cell Reverse Transcription-Polymerase Chain Reaction for the Simultaneous Detection of p210 and p190 BCR-ABL mRNAs in Chronic Myeloid Leukemia and Philadelphia-Positive Acute Lymphoblastic Leukemia Cell Lines
Articles in the same Issue
- The Basis of the Medicine of Tomorrow "Validating and Using Pharmacogenomics" Joint IFCC-Roche Diagnostics Conference, Kyoto, Japan, 1619 April 2000
- Diagnostics and the Future of Medicine
- Operomics: Molecular Analysis of Tissues from DNA to RNA to Protein
- Idiosyncratic Reactions to Drugs: Can Medicine Response Profiles Provide a Dynamic Drug Surveillance System?
- Hunting for Disease Genes in Multi-Functional Diseases
- Familial Studies on the Genetics of Cardiovascular Diseases: the Stanislas Cohort
- Quantitative PCR
- Gene Amplification as Means for Determining Therapeutic Strategies in Human Cancers
- Apolipoprotein E Polymorphisms and Concentration in Chronic Diseases and Drug Responses
- Angiotensin I-Converting Enzyme Gene Polymorphism and Drug Response
- Drug-Metabolizing Enzymes, Polymorphisms and Interindividual Response to Environmental Toxicants
- Database Analysis and Gene Discovery in Pharmacogenetics
- How to Manage Individualized Drug Therapy: Application of Pharmacogenetic Knowledge of Drug Metabolism and Transport
- P-Glycoprotein and Bioavailability-Implication of Polymorphism
- Cancer Therapy and Polymorphisms of Cytochromes P450
- Polymorphisms in UDP Glucuronosyltransferase Genes: Functional Consequences and Clinical Relevance
- The Human Multidrug Resistance-Associated Protein (MRP) Gene Family: From Biological Function to Drug Molecular Design
- Ethnic Differences in Drug Metabolism
- Hypervariable Region 1 of Hepatitis C Virus Genome and Response to Interferon Therapy
- A Functional Genomic Study of the Effects of Antipsychotic Agent Chlorpromazine in PC12 Cells
- Influence of Glutathione S-Transferase M1 and T1 Genotypes on Larynx Cancer Risk among Korean Smokers
- CYP2D6 Genotyping in Patients on Psychoactive Drug Therapy
- Genotyping of CYP2D6 in Parkinsons's Disease
- Rapid Analysis of CGG Repeat Length in the FMR1 Gene
- Multiplex In-cell Reverse Transcription-Polymerase Chain Reaction for the Simultaneous Detection of p210 and p190 BCR-ABL mRNAs in Chronic Myeloid Leukemia and Philadelphia-Positive Acute Lymphoblastic Leukemia Cell Lines
