Mycoplasma Protocols / Edition 1

by Roger J. Miles, Robin A. J. Nicholas

Overview

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 livesk 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 Prools 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.

Product Details

ISBN-13:	9780896035256
Publisher:	Springer-Verlag New York, LLC
Publication date:	05/25/1998
Series:	Methods in Molecular Biology , #104
Edition description:	1998
Pages:	330
Product dimensions:	6.10(w) x 9.25(h) x 0.04(d)

About the Author

Miles, Roger (King's Coll, London); Nicholas, Robin

Table of Contents

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.- Immunohishemical 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.

Editorial Reviews

Reviewer: Charles E. Edmiston, Jr., PhD (Medical College of Wisconsin)
Description: The mycoplasmas are a poorly understood but clinically significant group of pathogens causing disease in both the human and veterinary medicine populations. This book is a rather comprehensive effort to incorporate in a single text selected protocols encompassing isolation, biochemical characterization, diagnostic identification, and techniques to identify and manipulate the expression of target genes. It also discusses current techniques for identifying antigenic surface proteins.
Purpose: It presents in a concise, organized manner published protocols for the detection, isolation, identification, characterization, and genetic manipulation of Mycoplasma of both human and veterinary medicine interest.
Audience: This manual would be a valuable resource for either academic or industrial investigators whose area of study includes the pathogenic mycoplasma. Graduate students would also find this to be a valuable reference text for protocol development.
Features: The manual includes protocols for recovery of mycoplasma from humans and other animals and identification methods using traditional biochemicals, enzyme analysis, DNA probes, and immunologic assays. The editors have also included several chapters on techniques for transformation and transposon mutagenesis. In addition, several chapters describe the separation and characterization of membrane proteins which are key both for the understanding of the mechanisms of mycoplasma adherence to host cell surfaces and to the development of animal and human vaccines.
Assessment: While this is an extraordinarily ambitious text overall, the editors, with contributions from an outstanding group of internationally respected investigators, have succeeded in developing a superb manual that will have broad application in medical/veterinary diagnostic and research laboratories. This is an important text and should be part of the personal library of any investigator or graduate student working in this field of study.

Doody's Review Service

3 Stars from Doody

The mycoplasmas are a poorly understood but clinically significant group of pathogens causing disease in both the human and veterinary medicine populations. This book is a rather comprehensive effort to incorporate in a single text selected protocols encompassing isolation, biochemical characterization, diagnostic identification, and techniques to identify and manipulate the expression of target genes. It also discusses current techniques for identifying antigenic surface proteins. It presents in a concise, organized manner published protocols for the detection, isolation, identification, characterization, and genetic manipulation of Mycoplasma of both human and veterinary medicine interest. This manual would be a valuable resource for either academic or industrial investigators whose area of study includes the pathogenic mycoplasma. Graduate students would also find this to be a valuable reference text for protocol development. The manual includes protocols for recovery of mycoplasma from humans and other animals and identification methods using traditional biochemicals, enzyme analysis, DNA probes, and immunologic assays. The editors have also included several chapters on techniques for transformation and transposon mutagenesis. In addition, several chapters describe the separation and characterization of membrane proteins which are key both for the understanding of the mechanisms of mycoplasma adherence to host cell surfaces and to the development of animal and human vaccines. While this is an extraordinarily ambitious text overall, the editors, with contributions from an outstanding group of internationally respected investigators, have succeeded in developing a superb manualthat will have broad application in medical/veterinary diagnostic and research laboratories. This is an important text and should be part of the personal library of any investigator or graduate student working in this field of study.

Charles E. Edmiston

The 33 contributions, written by active laboratory-based research workers, provide methods for the detection, isolation, identification, and characterization of mycoplasmas, with an emphasis on those of medical and veterinary significance. Coverage includes biochemical, genetic, and molecular techniques essential to understanding pathogenicity and adhesion to host cells, as well as protocols for the detection of mycoplasma contamination in cell cultures. Annotation c. by Book News, Inc., Portland, Or.

Booknews

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

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