Microbiological Assay

Microbiological Assay

An Introduction to quantitative principles and Evaluation

1st Edition - January 28, 1977

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  • Author: William Hewitt
  • eBook ISBN: 9780323157094

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Microbiological Assay: An Introduction to Quantitative Principles and Evaluation aims to provide an introduction to the principles of microbiological assay, assay design, and calculation procedures. Organized into nine chapters, this book begins with the philosophy of biological assay, as well as the method’s basic techniques, principles, mechanization, automation, purpose, reference standards, specifications, and reports. It also looks into the preparation of test solutions of standard and sample. Some chapters follow explaining the specific methods, such as agar diffusion assay and tube assay; others explore the statistical evaluation of these assays. Features of assay design, such as replication, number of dose levels, and spacing of dose levels, are also described. This book will serve as an elementary introduction to this field of interest to help encourage a less empirical approach to the subject.

Table of Contents

  • Preface


    Chapter 1 Introduction

    1.1 Philosophy of Biological Assay

    1.2 Basic Techniques and Principles

    1.3 Mechanization and Automation

    1.4 Purpose of the Assay

    1.5 Reference Standards

    1.6 Preparation of Test Solutions of Standard and Sample

    1.7 Specifications and Reports


    Chapter 2 The Agar Diffusion Assay

    2.1 Introduction

    2.2 Theory of Zone Formation

    2.3 Nature of the Response Curve

    2.4 Dose-Response Curves in Practice

    2.5 Simple Assay Designs

    2.6 Simple Multiple Assay Designs

    2.7 Simplification of Computation of the Potency Ratio

    2.8 Designs Incorporating Checks for Curvature (Three Dose Levels)

    2.9 Designs Incorporating Checks for Curvature (Four Dose Levels)

    2.10 Assays by Large Plates—General Principles

    2.11 Large Plate Assays Using Latin Square Designs

    2.12 Large Plate Assays Using Quasi-Latin Square Designs

    2.13 Low-Precision Assays Using Large Plates

    2.14 Small Plate Assays Using Interpolation from a Standard Curve

    2.15 Missing Values


    Chapter 3 Tube Assays Growth-Promoting Substances

    3.1 General Principles

    3.2 Measurement of Response

    3.3 Nonideal Responses

    3.4 Slope Ratio Assays—An Unbalanced Design

    3.5 Criteria of Validity

    3.6 Multiple Linear Regression Equations

    3.7 Potency Computation from an Unbalanced Design

    3.8 Balanced Slope Ratio Assays with Linearity Check

    3.9 Simplified Computation of Potency Ratio from Balanced Assays

    3.10 Multiple Assays by the Slope Ratio Method


    Chapter 4 Tube Assays for Antibiotics

    4.1 General Principles

    4.2 Response Curve—Commonly Used Forms of Expression

    4.3 Linearization of Dose-Response Relationships—Theoretical Considerations

    4.4 Dose-Response Linearization Procedures in Practice

    4.5 Potency Estimation by Interpolation from a Standard Curve

    4.6 Graphic Estimation of Potency by Probit of Response versus Dose

    4.7 Arithmetical Estimation of Potency from an Assay of Balanced Design by Angular Transformation of Response

    4.8 Graphic Estimation of Potency Using the Relationship Log Response versus Dose

    4.9 Estimation of Potency by Interpolation from a Dose-Response Line Using an Automated System


    Chapter 5 Assay of mixtures of Antibiotics

    5.1 Occurrence of Mixtures and the Nature of the Problem

    5.2 General Techniques

    5.3 Comparative Bioautographs

    5.4 Quantitative Bioautographs

    5.5 Differential Assays


    Chapter 6 Evaluation of Parallel Line Assays

    6.1 Introduction

    6.2 Basic Assumptions for the Statistical Evaluation of Parallel Line Assays

    6.3 Evaluation of a Standard Log Dose-Response Curve

    6.4 Analysis of Variance to Separate Components Attributable to Various Sources

    6.5 Evaluation of a Simple Two Dose Level Assay

    6.6 An Alternative Method for Obtaining Confidence Limits

    6.7 Evaluation of a Multiple Two Dose Level Assay

    6.8 Evaluation of a Three Dose Level Assay

    6.9 Evaluation of a Large Plate Assay (Latin Square Design)

    6.10 Evaluation of a Large Plate Assay (Quasi-Latin Square Design)

    6.11 Evaluation of a Large Plate, Low Precision Assay

    6.12 Evaluation of an Assay Incorporating Reference Points—The "FDA" Design

    6.13 Evaluation of a Quantitative Bioautograph

    6.14 Evaluation of Tube Assays Using Function of Response versus Logarithm of Dose


    Chapter 7 Evaluation of Slope Ratio Assays

    7.1 Principles of Evaluation

    7.2 Evaluation of Simple Slope Ratio Assays

    7.3 Evaluation of a Multiple Slope Ratio Assay

    7.4 Evaluation of a Turbidimetric Antibiotic Assay by Angular Transformation of Response and Slope Ratio


    Chapter 8 Choice of Method and Design

    8.1 Choice of Method

    8.2 General Considerations in Selection of Design

    8.3 Plate Assay Designs

    8.4 The Influence of Curvature in Parallel Line Assays

    8.5 Choice of Design for Plate Assay

    8.6 Tube Assays—General Considerations

    8.7 Designs for Slope Ratio Assays

    8.8 Influence of Curvature in Slope Ratio Assays

    8.9 Choice of Design for Slope Ratio Assays

    8.10 Tube Assays for Antibiotics


    Chapter 9 Repeated Assays, Specifications, and Reports

    9.1 Replication of Assays

    9.2 Collaborative Assays

    9.3 Combination of Replicate Potency Estimates

    9.4 Combination of Replicate Potency Estimates—Simplified Methods

    9.5 Specifications for Antibiotics

    9.6 Official Standards—A Practical Approach


    Appendix 1 Patterns for Small Plate Assays

    Appendix 2 Calculations for Parallel Line Assays

    Appendix 3 Potency Ratio, or F/E, Tables

    Appendix 4 Proforma and Worked Example—Agar Diffusion Assay

    Appendix 5 Sources of Reference Materials

    A5.1 International Biological Standards and International Biological Reference Preparations

    A5.2 British Biological Standards and Reference Preparations

    A5.3 European Pharmacopeia Commission Reference Substances

    A5.4 United States Pharmacopeia Reference Substances

    A5.5 International Chemical Reference Substances

    Appendix 6 The Dilution Of Reference Standards

    Appendix 7 The Probit Transformation

    Appendix 8 The Angular Transformation

    Appendix 9 The t Distribution

    Appendix 10 Variance Ratio Tables—The F Test

    Appendix 11 The χ2 Distribution

    Appendix 12 The Range/Mean Test

    Appendix 13 Evidence That Quadratic Curvature is Without Influence in Balanced Parallel Line Assays


Product details

  • No. of pages: 298
  • Language: English
  • Copyright: © Academic Press 1977
  • Published: January 28, 1977
  • Imprint: Academic Press
  • eBook ISBN: 9780323157094

About the Author

William Hewitt

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