Mycoplasma Protocols

ISBN-13: 9781617370618 / Angielski / Miękka / 2010 / 330 str.

Roger J. Miles; Robin A. J. Nicholas

Opis

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.

Recenzje

"The value of this book lies in the amount of technical detail provided. . .this should enable the interested investigator to carry out the protocols described without recourse to other publications."-Journal of Medical Microbiology

Spis treści

Introduction R. Miles and R. Nicholas The Medical Significance of Mycoplasmas P. Taylor The Veterinary Significance of Mycoplasmas R. Nicholas Recovery of Mycoplasmas from Man P. Taylor Recovery of Mycoplamas from Animals R. Nicholas and S. Baker Recovery of Mycoplasmas from Birds J. Bradbury Cultivation of Ureaplasmas P. Furr Quality Control Testing of Mycoplasma Media D. Windsor and H. Windsor Biochemical Characteristics of Mycoplasma Identification J. Poveda Enzyme Analysis: The Rationale and Use of Enzyme Assays in Assigning Function to Gene Nucleotide Sequences and the Procedures for the Assay of Three Enzymatic Functions Conserved in All Mollicutes J. D. Pollack Determination of Substrate Utilization Rates by Mycoplasmas R. Miles and C. Agbanyim Serological Identification of Mycoplasmas by Growth and Metabolism Inhibition Tests J. Poveda and R. Nicholas Identification of Mycoplasmas by Dot Immunobinding on Membrane Filtration (MF dot) F. Poumarat Identification of Mycoplasmas by Immunofluorescence J. Bradbury Diagnostic Application of Monoclonal Antibody-Based Sandwich ELISAs H. Ball and D. Finlay Immunocytochemical staining of Fixed Tissues E. Scanziani Extraction of DNA from Mycoplasmas J. Bashiruddin Characterization of Mycoplasmas by PCR and Sequence Analysis with Universal 16S rDNA Primers K. E. Johansson, M. Heltander, and B. Petterson PCR and RFLP Methods for Specific Detection and Identification of Mycoplasma Mycoides Subsp. Mycoides SC J. Bashiruddin Characterization of Mycoplasmas by RAPD Fingerprinting G. Rawadi Classification of Isolates by DNA-DNA Hybridization K. Sachse and H. Hotzel. Insertion Sequence Analysis J. Frey Detection of Mycoplasmas in Cell Cultures by Cultural Methods G. Masover and F. Becker Detection of Mycoplasmas in Cell Culture by Fluorescence Methods G. Masover and F. Becker Transformation of Mycoplasmas F. Chris Minion, and P. Kapke Transposon Mutagenesis L. Voelker and K. Dybvig Demonstration of Extrachromosomal Elements L. Voelker and K. Dybvig Expression of Foreign Genes in Acholeplasma T. Jarhede and A. Wieslander Mycoplasma Gene Expression in Escherichia Coli F. C. Minion Polyacrylamide Gel Electrophoresis Separation of Whole Cell Proteins M. Duffy, A. Noormohammadi, N. Baseggio, G. Browning, and P. Markham Immunological and Biochemical Characterization of Membrane Proteins M. Duffy A. Noormohammadi N. Baseggio, G. Browning, and P. Markham Detection and Analysis of Mycoplasma Adhesins K. Sachse Transmission Electron Microscopy and Immunogold Staining of Mycoplasma Surface Antigens G. Christiansen and S. Birkelund Index.

Blurb

In Mycoplasma Protocols, Roger Miles and Robin Nicholas present a collection of cutting-edge 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 pathogenicity 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.

Mycoplasma Protocols provides up-to-date and easy-to-follow mycoplasma methods for practical application in medical and veterinary diagnostic and research laboratories. The techniques permit effective work with these normally fastidious microorganisms, allowing investigators to illuminate their roles across a wide range of chronic respiratory, arthritic, and urogenital diseases. For all those working in this important field, Mycoplasma Protocols immediately becomes the methodological resource of choice.