Methods in Mycoplasmology V1

Methods in Mycoplasmology V1

Mycoplasma Characterization

1st Edition - January 28, 1983

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  • Editor: Samuel Razin
  • eBook ISBN: 9780323147132

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Description

Methods in Mycoplasmology, Volume I: Mycoplasma Characterization is the first of a two-volume series that resulted from cumulative efforts to meet the need for standardized techniques in mycoplasmology. The book is organized into five sections. Section A provides an introduction to the mycoplasmas by discussing their characteristics and classification, mycoplasma-animal host interrelationships, and mycoplasma-plant-insect interrelationships. The chapters in Section B examine the morphological and ultrastructural features of mycoplasmas, using techniques such as phase-contrast microscopy, dark-field microscopy, and transmission electron microscopy. Section C discusses the cultivation and nutrition of mycoplasmas. Section D focuses on analytical methods for the characterization of mycoplasmas, including harvest and washing procedures, cell lysis and membrane isolation, and crossed immunoelectrophoresis. Section E describes tests designed to detect specific enzymatic activities or nutritional requirements of mycoplasmas. These include sugar fermentation and arginine hydrolysis tests as well as tests to establish the sterol requirement of mycoplasmas. Section F deals with serological tests. These tests either have specific application to detection of mycoplasma antigens or antibody in plant and animal tissues; are useful in identification of specific groups of mycoplasmas (spiroplasmas); or provide higher sensitivity in measurements of antibodies against mycoplasmas.

Table of Contents


  • Contributors

    Preface

    Contents of Volume II

    Section A Introduction

    A1 Characteristics of the Mycoplasmas as a Group

    A2 Principles of Mycoplasma Classification and Taxonomy

    A3 Mycoplasma–Animal Host Interrelationships

    A4 Mycoplasma–Plant–lnsect Interrelationships

    Section B Morphology and Ultrastructure

    B1 Introductory Comments

    B2 Phase-Contrast Microscopy

    B3 Dark-Field Microscopy

    B4 Gram Staining Technique

    B5 Transmission Electron Microscopy: Basic Techniques

    B6 Scanning Electron Microscopy of Mycoplasmas

    B7 Specialized Electron Microscopic Techniques for Mycoplasmas in Animal Tissues

    B8 Specialized Electron Microscopic Techniques for Mycoplasma-Like Organisms in Plant Tissues

    B9 Specialized Electron Microscopic Techniques for Spiroplasmas in Plant and Insect Tissues

    B10 Transfer of Spiroplasmas onto Collodion Membranes for Electron Microscopic Studies

    B11 Identification of Mycoplasma Colonies

    B12 Examination of Mycoplasmas for Motility

    Section C Cultivation and Nutrition

    C1 Introductory Remarks

    C2 General Cultivation Techniques for Mycoplasmas and Spiroplasmas

    C3 Collection and Transport of Specimens

    C4 Growth Inhibitory Factors in Animal and Plant Tissues

    C5 Precautions in Handling Infectious Material

    C6 Sterility and Quality Control of Mycoplasma Culture Media

    C7 Culture Media for Classical Mycoplasmas

    C8 Culture Media for Ureaplasmas

    C9 Culture Media for Spiroplasmas

    C10 Culture Media for Anaeroplasmas

    C11 Defined and Partly Defined Media

    C12 Cloning and Filtration Techniques for Mycoplasmas

    C13 Labeling of Cellular Components during Growth

    C14 Methods for Direct and Indirect Measurement of Mycoplasma Growth

    C15 Preservation of Mycoplasma Cultures and Culture Collections

    C16 Bacterial and Fungal Inhibitors in Mycoplasma Culture Media

    C17 Optimal Temperature and Atmospheric Conditions for Growth

    Section D Analysis of Cellular Components

    D1 Introductory Remarks

    D2 Harvest and Washing of Mycoplasmas

    D3 Cell Lysis and Isolation of Membranes

    D4 Manipulation of Membrane Lipid Composition

    D5 Electrophoretic Characterization of Mycoplasma Proteins

    D6 Characterization of Membrane Proteins by Crossed Immunoelectrophoresis

    D7 Characterization of Membrane Lipids

    D8 Characterization of Membrane Lipoglycans

    D9 Characterization of Carbohydrate Components of Mycoplasma Membranes

    D10 DNA Extraction and Purification

    D11 Determination of Guanine Plus Cytosine Content of DNA

    D12 Genome Size Determination

    D13 EcoRI Restriction Enzyme Analysis of Mycoplasma DNA

    D14 Hybridation between Mycoplasma DNAs

    D15 Localization of Enzymes in Mycoplasmas: Preparatory Steps

    Section E Biochemical and Enzymatic Tests in Mycoplasma Identification

    E1 Introductory Remarks

    E2 Sugar Fermentation

    E3 Arginine Hydrolysis

    E4 Urea Hydrolysis

    E5 Tests for Digitonin Sensitivity and Sterol Requirement

    E6 Phosphatase Activity

    E7 Proteolytic Activity

    E8 Film and Spot Production

    E9 Pigmented Carotenoids

    E10 Tetrazolium Reduction

    E11 Hemagglutination, Hemadsorption, and Hemolysis

    E12 Detection of β-D-Glucosidase: Hydrolysis of Esculin and Arbutin

    E13 Isozyme Resolution in Mycoplasmas

    Section F Serological Identification of Mycoplasmas

    F1 Introductory Remarks

    F2 Preparation of Antigens and Antisera

    F3 Growth Inhibition Tests

    F4 Metabolism Inhibition Tests

    F5 Tetrazolium Reduction Inhibition

    F6 The Mycoplasmacidal Test (MCT)

    F7 Immunoflourescence

    F8 Agar Precipitin and Immunoelectrophoretic Methods for Detection of Mycoplasmic Antigens

    F9 Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Anti-Mycoplasmal Antibody

    F10 Application of ELISA to Spiroplasma Detection and Classification

    F11 The Combined Deformation-Metabolism Inhibition Test

    F12 Single Radial Hemolysis Technique

    F13 Growth Precipitation Test

    Appendix: Tables 1-4

    Index


Product details

  • No. of pages: 526
  • Language: English
  • Copyright: © Academic Press 1983
  • Published: January 28, 1983
  • Imprint: Academic Press
  • eBook ISBN: 9780323147132

About the Editor

Samuel Razin

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