PCR Topics

PCR, developed at Cetus Corporation/USA by Henry A. Erlich, Kary Mullis and Randall K. Saiki, is a very simple method for amplifying nucleic acids in vitro. The realization of this idea bases on the repetition of a set of three different temperatures and yields an increase of the target structure up to a factor of 106 to 1012. Therefore, this technique is predisposed for safe analysis and characterization of DNA and RNA sequences of interest, even where the starting amount of material is enormously small. Because of its sensitivity, speed and versatility this method is particularly suitable for investigations of oncogenes, tumor associated translocations, retroviral sequences, lymphokines and mainly the broad field of degenerative and inflammatory diseases of nervous system. PCR seems to be the technique which could overcome the two most important problems in that field: very small amount of material combined with the necessity of rapid diagnostic procedures in inflammatory infections. "PCR topics" will give an actual overview of basic and applied research fields on usage of polymerase chain reaction. All contributions to this book have been presented at an international congress on "Usage of Polymerase chain reaction in genetic and infectious diseases" which took place in june 1990 in Berlin. The editors wish to thank all participants for their contributions. We offer our thanks and gratitude to our coworkers and especially to our technical assistents Barbara Trampenau, Mirjana Wiirdemann and Hannelore Leonhard.

Basic Methodology and Research Applications. - Quantitation of mRNA by the Polymerase Chain Reaction. - Sequencing of PCR Products Analysis of Factor IX Genes and of Recombination Events in Immunoglobulin Genes. - Alternative Methods for DNA Probing in Diagnosis: Ligase Chain Reaction (LCR). - Application of PCR to the Detection and Analysis of Point Mutations in the Human Factor VIII Gene. - The Use of PCR in the Diagnosis of Muscle Disease. - Taq DNA Polymerase-Synthesized Single-Stranded DNA Hybridization Probes and their Application in Northern Blotting and in situ Hybridization. - Computer Program for the Selection of PCR Primers. - Nervous System Cross Reactivity Sequences. - The Use of Short Adapters for Priming PCR of Unknown Chromosomal DNA Fragments. - Labeling of Specific DNA Sequences With Digoxigenin during Polymerase Chain Reaction. - Quantitative Analysis of Polymerase Chain Reaction (PCR) Products by Means of Fluorescence Detection. - Reverse Transcription PCR (RT-PCR) of Parathyroid Hormone (PTH) mRNA. - False Positive Reactions in PCR. - Error Production and Error Propagation During PCR. - Genetics and Evolution. - PCR in Linkage Analysis of Genetic Diseases. - Amplification, Isolation and Direct Nucleotide Determination of Entire Genes: Application to the Study of 16S rRNAs for Molecular Evolution in Bacteria, Identification of Cultural Isolates and Development of Probes. - Diagnosis of Cystic Fibrosis Detection of the Gene Deletion Delta F 508. - Application of PCR for Prenatal Diagnosis of Cystic Fibrosis (CF). - The Use of Non-Viable Cultured Amniotic Fluid Cells for Prenatal Diagnosis of Cystic Fibrosis by Polymerase Chain Reaction. - Determination of Glutathione S-Transferase u Genotype in Humans by PCR-Amplified Gene-Probes. - Rapid Apolipoprotein E GenotypeIdentification. - Detection and Characterization of Viral Infections. - Herpes Viruses. - Detection of Herpes Viruses by PCR Diagnostic Aspects. - Diagnosis of Herpes Simplex Virus Encephalitis (HSVE) by Polymerase Chain Reaction. - Rapid Detection of CMV-Specific DNA and mRNA by PCR in Immunocompromised Patients. - Specific and Sensitive Detection of Cytomegalovirus DNA in Clinical Specimens by PCR. - Detection of EBNA2-mRNA in Primary Central Nervous System Lymphomas in Patients with AIDS. - Detection of Epstein-Barr Virus Genomes in Lymphoproliferative Diseases. - Screening of Lymphatic Tissues for the Presence of Human Herpesvirus 6 DNA Using the Polymerase Chain Reaction (PCR). - Retroviruses. - HTLV Retrovirus in Neurological Disease. - Use of PCR to Investigate the Cellular Tropism of Human T Cell Leukemia Virus Type I. - Viral Genome Sequences in Cerebrospinal Fluid and Serum of HIV-1 Infected but Asymptomatic Patients. - Hepatitis Viruses. - PCR Detection of HBV Replication in Anti-HBeAg Chronic Carriers with Pre-S Antigenemia. - Detection of Virus DNA by PCR in Hepatitis Non-A, Non-B. - Hepatitis B Virus DNA Identification by Polymerase Chain Reaction (PCR) and Correlation with Serological HBV Markers. - Detection and Characterization of Bacterial Infections. - Rapid Diagnosis of Mycobacterium Tuberculosis Infection by PCR. - Detection of Chlamydia Trachomatis by PCR. - The Polymerase Chain Reaction (PCR) to Detect Gene Sequences of Borrelia Burgdorferi, the Etiologic Agent of Lyme Disease. - Rapid Diagnosis of Human Listeriosis Using the Polymerase Chain Reaction. - Oncogenes. - Quantitation of Mdr1 Transcript by PCR a Tool for Monitoring Drug Resistance in Cancer Chemotherapy. - Ras Oncogene Mutations in Renal Cell Cancer. - Primer Mismatch Analysis: a Rapid and Non-IsotopicMethod for Analysing Ras Mutations in Human Cancer. - Expression of Adenovirus Type 12 13S Gene Product is Sufficient to Immortalize Primary Mouse Cells. - PCR-Based Rapid Detection of the Philadelphia-Chromosome in Chronic Myelogenous and Acute Lymphoblastic Leukemia. - List of Contributors.