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.
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
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.
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