PCR Topics : Usage of Polymerase Chain Reaction in Genetic and Infectious Diseases

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. Front Matter....Pages I-XI Front Matter....Pages 1-1 Quantitation of mRNA by the Polymerase Chain Reaction....Pages 3-8 Sequencing of PCR Products — Analysis of Factor IX Genes and of Recombination Events in Immunoglobulin Genes....Pages 9-14 Alternative Methods for DNA Probing in Diagnosis: Ligase Chain Reaction (LCR)....Pages 15-22 Application of PCR to the Detection and Analysis of Point Mutations in the Human Factor VIII Gene....Pages 23-31 The Use of PCR in the Diagnosis of Muscle Disease....Pages 32-40 Taq DNA Polymerase-Synthesized Single-Stranded DNA Hybridization Probes and their Application in Northern Blotting and in situ Hybridization....Pages 41-45 Computer Program for the Selection of PCR Primers....Pages 46-47 Nervous System Cross Reactivity Sequences....Pages 48-52 The Use of Short Adapters for Priming PCR of Unknown Chromosomal DNA Fragments....Pages 53-55 Labeling of Specific DNA Sequences with Digoxigenin during Polymerase Chain Reaction....Pages 56-58 Quantitative Analysis of Polymerase Chain Reaction (PCR) Products by Means of Fluorescence Detection....Pages 59-62 Reverse Transcription PCR (RT-PCR) of Parathyroid Hormone (PTH) mRNA....Pages 63-65 False Positive Reactions in PCR....Pages 66-68 Error Production and Error Propagation During PCR....Pages 69-71 Front Matter....Pages 73-73 PCR in Linkage Analysis of Genetic Diseases....Pages 75-79 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....Pages 80-83 Diagnosis of Cystic Fibrosis — Detection of the Gene Deletion Delta F 508....Pages 84-88 Application of PCR for Prenatal Diagnosis of Cystic Fibrosis (CF)....Pages 89-91 The Use of Non-Viable Cultured Amniotic Fluid Cells for Prenatal Diagnosis of Cystic Fibrosis by Polymerase Chain Reaction....Pages 92-93 Determination of Glutathione S-Transferase u Genotype in Humans by PCR-Amplified Gene-Probes....Pages 94-98 Front Matter....Pages 73-73 Rapid Apolipoprotein E Genotype Identification....Pages 99-102 Front Matter....Pages 103-103 Detection of Herpes Viruses by PCR-Diagnostic Aspects....Pages 105-112 Diagnosis of Herpes Simplex Virus Encephalitis (HSVE) by Polymerase Chain Reaction....Pages 113-116 Rapid Detection of CMV-Specific DNA and mRNA by PCR in Immunocompromised Patients....Pages 117-123 Specific and Sensitive Detection of Cytomegalovirus DNA in Clinical Specimens by Polymerase Chain Reaction....Pages 124-129 Detection of EBNA2-mRNA in Primary Central Nervous System Lymphomas in Patients with AIDS....Pages 130-135 Detection of Epstein-Barr Virus Genomes in Lymphoproliferative Diseases....Pages 136-141 Screening of Lymphatic Tissues for the Presence of Human Herpesvirus 6 DNA Using the Polymerase Chain Reaction (PCR)....Pages 142-144 HTLV Retrovirus in Neurological Disease....Pages 147-160 Use of PCR to Investigate the Cellular Tropism of Human T Cell Leukemia Virus Type 1....Pages 161-170 Viral Genome Sequences in Cerebrospinal Fluid and Serum of HIV-I-Infected but Asymptomatic Patients....Pages 171-177 PCR Detection of HBV Replication in Anti-HBeAg Chronic Carriers with Pre-S Antigenemia....Pages 181-185 Detection of Virus DNA by PCR in Hepatitis Non-A, Non-B....Pages 186-191 Hepatitis B Virus DNA — Identification by Polymerase Chain Reaction (PCR) and Correlation with Serological HBV Markers....Pages 192-195 Front Matter....Pages 197-197 Rapid Diagnosis of Mycobacterium Tuberculosis Infection by PCR....Pages 199-202 Detection of Chlamydia Trachomatis by PCR ....Pages 203-205 The Polymerase Chain Reaction (PCR) to Detect Gene Sequences of Borrelia Burgdorferi, the Etiologic Agent of Lyme Disease....Pages 206-208 Rapid Diagnosis of Human Listeriosis Using the Polymerase Chain Reaction....Pages 209-213 Front Matter....Pages 215-215 Quantitation of Mdr1 Transcript by PCR a Tool for Monitoring Drug Resistance in Cancer Chemotherapy....Pages 217-220 Ras Oncogene Mutations in Renal Cell Cancer....Pages 221-223 Front Matter....Pages 215-215 Primer Mismatch Analysis: a Rapid and Non-Isotopic Method for Analysing Ras Mutations in Human Cancer....Pages 224-229 Expression of Adenovirus Type 12 13S Gene Product is Sufficient to Immortalize Primary Mouse Cells....Pages 230-233 PCR-Based Rapid Detection of the Philadelphia-Chromosome in Chronic Myelogenous and Acute Lymphoblastic Leukemia....Pages 234-238 Back Matter....Pages 239-258 The Polymerase chain reaction (PCR) has revolutionized the technical development of molecular biology and immunology. It is a very simple method for amplifying nucleic acids in vitro. During the last two years PCR has mostly been used on investigations of oncogenes, tumor associated translocations, retroviral structures, HLA polymorphism and lymphokines. Because this method will make it possible to identify previously unknown structures it is particulary suitable for diagnosis in the broad field of degenerative and inflammatory diseases of the nervous system. Here we have two problems: a small amount of material and the necessity of rapid diagnostic procedures in inflammatory infections. PCR seems to be the technique which could overcome these difficulties