Nucleic Acid Probes In Diagnosis Of Human Genetic Diseases

Reviews the basic terminology and techniques of nucleic acid probes, how such probes can be used in the detection and analysis of human genetic disorders, and the impact of this technology on public health care services and on affected individuals and their families. Addresses use of nucleic acid probes in prenatal and carrier detection of Duchenne muscular dystrophy, the cystic fibrosis region of chromosome 7, and the fragile X chromosome. Also discusses ethical issues raised by use of this technology, the role of regulatory agencies, and the establishment of a centralized DNA-based genetic reference laboratory.

With each chapter written by a distinguished expert in the biochemistry field, this comprehensive volume describes the preparation and use of a variety of radioactive and non-radioactive probes in situations ranging from research laboratories to routine diagnosis laboratories. The enzymatic and chemical techniques for labeling nucleic acid probes with radioisotopes and with non-radioactive ligands and haptens are discussed. Additionally, the associated methods for their detection, the use of these probes in the diagnosis of human and microbial pathogens, of plant viruses and viroids, and of human genetic disorders, as well as in the detection of nucleic acids in tissues and cells by in situ hybridization are presented.

The aim of molecular diagnostics is preferentially to detect a developing disease before any symptoms appear. There has been a significant increase, fueled by technologies from the human genome project, in the availability of nucleic acid sequence information for all living organisms including bacteria and viruses. When combined with a different type of instrumentation applied, the resulting diagnostics is specific and sensitive. Nucleic acid-based medical diagnosis detects specific DNAs or RNAs from the infecting organism or virus and a specific gene or the expression of a gene associated with a disease. Nucleic acid approaches also stimulate a basic science by opening lines of inquiry that will lead to greater understanding of the molecules at the center of life. One can follow Richard Feynman’s famous statement “What I cannot create, I do not understand.”

Nucleic Acid Testing for Human Disease describes various techniques including target and signal amplification-based NAT procedures, microarrays, bead-based multiplex assays, in situ hybridization, and SNP techniques. This book discusses RNA expression profiling and laboratory issues such as the need for proper validation of tests intended fo

Senior scientists Marilena Aquino de Muro and Ralph Rapley have brought together an outstanding collection of time-tested protocols for designing and using genes probes in a wide variety of applications. The applications covered range from searching for specific genes in the human chromosome to the detection of microorganisms and their toxins in the environment and in food samples, as well as in the diagnosis of human disease. Helpful tutorials explain the principles of gene probe design, labeling, detection, target formation, and hybridization conditions.

This book covers all the commonly used methods for the analysis and diagnosis of human genetic diseases. The chapters cover the analysis of gene mutations as well as the functional organization of genes and chromosomal regions. The techniques for the analysis of human genetic disease have advanced so rapidly in recent years that almost all the chapters in this new edition have significant additions. For example, when the first edition of this book was published, prenatal diagnosis based on DNA analysis was performed in two weeks. The advent of PCR has changed this to just a few hours and has had an impact in almost all areas of molecular medicine. The expanded chapters now have sections also covering: fluorescence in situ hybridization (FISH), synthetic oligonucleotide hybridization, and pulsed-field gel electrophoresis. This book should be a valuable text for scientists working in a pure research environment as well as those in service laboratories analyzing different human disease mutations.

Nucleic acid hybridizations: principles and strategies; Generation of nucleic acid probe molecules; Nonisotopic DNA labeling strategies; Approaches to genetic analysis: genetic disease and screening; Approaches to genetic analysis: forensics; Approaches to infectious disease diagnosis; Diagnostic nucleic acid probe technology in veterinary medicine; Diagnostic nucleic acid probes in human medicine: applications, present and future; Developments in food technology: applications and regulatory and economic considerations; Current development and applications of nucleic acid technology in the environmental sciences.