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Methods in Mycoplasmology V1 - 1st Edition - ISBN: 9780125838016, 9780323147132

Methods in Mycoplasmology V1

1st Edition

Mycoplasma Characterization

Editor: Samuel Razin
eBook ISBN: 9780323147132
Imprint: Academic Press
Published Date: 28th January 1983
Page Count: 526
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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



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



No. of pages:
© Academic Press 1983
28th January 1983
Academic Press
eBook ISBN:

About the Editor

Samuel Razin

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