Mycoplasma Protocols

Over the last decade, interest in mycoplasmas has been greatly sti- lated by the spread of contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides SC, in Europe and Africa and the discovery of a possible association between mycoplasmas and AIDS. During this period there has also been a recognition by control authorities and int- national organizations, including the Office International des Epizooties, that mycoplasmas are major causes of economic loss in livestock and continue to be a problem in humans as a result of a range of chronic respiratory and urogenital diseases. However, it is likely that the true significance of my- plasmas is still underestimated because of the difficulties in working with these extremely fastidious microorganisms. Advances in such laboratory technology as DNA amplification has enabled the detection of unrecoverable mycoplasmas in tissue samples and this will provide us with a better understanding of their role in disease. Important developments have also been seen in more conv- tional areas such as serological tests and media formulation enabling improved recovery and identification. Mycoplasma Protocols provides in a single volume up-to-date and easy-- follow methods for the detection, isolation, identification, and characterization of mycoplasmas, with a major emphasis on those of medical and veterinary significance. It also includes biochemical, genetic, and molecular techniques that will form the basis of understanding pathogenicity and, particularly, - hesion to host cells.

Klappentext

This is a collection of cutting-edge mycoplasma methods for the detection, isolation, identification, characterization, and genetic manipulation of the pathogenic mycoplasmas. These step-by-step methods are crafted for successful reproducibility and include biochemical, genetic, and molecular techniques essential to understanding pathogenecity and adhesion to host cells. They also cover the detection of mycoplasmas in cell cultures, an important tool not only in viral diagnosis and research, but also in the production of vaccines and various biological products.

Inhalt
Medical Significance of Mycoplasmas.- The Veterinary Significance of Mycoplasmas.- Recovery of Human Mycoplasmas.- Recovery of Mycoplasmas from Animals.- Recovery of Mycoplasmas from Birds.- Cultivation of Ureaplasmas.- Quality Control Testing of Mycoplasma Medium.- Biochemical Characteristics in Mycoplasma Identification.- Enzyme Analysis.- Determination of Substrate Utilization Rates by Mycoplasmas.- Serological Identification of Mycoplasmas by Growth and Metabolism Inhibition Tests.- Identification of Mycoplasmas by Dot Immunobinding on Membrane Filtration (MF Dot).- Identification of Mycoplasmas by Immunofluorescence.- Diagnostic Application of Monoclonal Antibody (MAb)-Based Sandwich ELISAs.- Immunohistochemical Staining of Fixed Tissues.- Extraction of DNA from Mycoplasmas.- Characterization of Mycoplasmas by PCR and Sequence Analysis with Universal 16S rDNA Primers.- PCR and RFLP Methods for the Specific Detection and Identification of Mycoplasma mycoides subsp. mycoides SC.- Characterization of Mycoplasmas by RAPD Fingerprinting.- Classification of Isolates by DNA-DNA Hybridization.- Insertion Sequence Analysis.- Detection of Mycoplasmas in Cell Cultures by Cultural Methods.- Detection of Mycoplasmas in Cell Culture by Fluorescence Methods.- Transformation of Mycoplasmas.- Transposon Mutagenesis.- Demonstration of Extrachromosomal Elements.- Expression of Foreign Genes in Acholeplasma laidlawii.- Mycoplasma Gene Expression in Escherichia coli.- Polyacrylamide Gel-Electrophoresis Separation of Whole-Cell Proteins.- Immunological and Biochemical Characterization of Membrane Proteins.- Detection and Analysis of Mycoplasma Adhesins.- Transmission Electron Microscopy and Immunogold Staining of Mollicute Surface Antigens.