Physical Methods for Microorganisms Detection

W. H. Nelson

• Publisher: CRC Press
• Year: 2018
• Print ISBN: 9781315896496
• eBook ISBN: 9781351092494
• Edition: 1
• Topic: Ciencias biológicas
• Subtopic: Biología

This volume presents detection and identification methods for bacteria and yeast. Chapters are written by expert laboratory practitioners and instrument makers and focuses on those methods that show widespread practical application, such as ATP luminescence. Food applications include rapid detection and quantitation of bacteria in raw milk, pasteurized milk, other dairy products, and raw meat. Other topics include brewing applications for beverages, starter culture monitoring, clinical analyses, blood and urine analysis procedures, analysis of aerosols, bioprocess safety, and biodeterioration. This book is a must for microbiologists in food quality labs and clinical labs.

Chapter 1
Bioluminescence in Clinical Microbiology

Charles T. Gregg

Table of Contents

I. Introduction

A. What is Bioluminescence?

B. Mechanism Studies, Properties of Bioluminescence

II. Bioluminescence as a Rapid Screen for Urinary Tract Infection (UTl)

A. Early Work

B. The Sociology of UTI

C. Techniques for Urine Screening

D. Criteria for Evaluation of Urine Screens

E. Future Developments

III. Bioluminescence for Rapid Antibiotic Susceptibility Testing (AST)

A. Introduction

B. Early Work

C. Current Standard Methods

D. The Bioluminescence AST

E. The LAD Sensi-Quik®

1. Introduction

2. Medium Studies

3. Ionic Strength Effects

4. Incubation Times

5. Antibiotic Concentration Curves

6. Error Rates

7. Microtiter Plate (MTP) Luminometer Evaluations

8. Current Status

IV. Concluding Remarks

Acknowledgments

References

Then shall thy light spring forth as the morning, and thine health shall spring forth speedily.

Isaiah 58:8

1. Introduction

A. What is Bioluminescence?

The quote from Isaiah succinctly summarizes the concerns of this chapter, i.e., light (from bioluminescence) and its role in health care.

Luminescence is a term coined in 1888 by the physicist Eilert Weidemann to distinguish "cold" light from the light arising from incandescence.¹'² Weidemann indicated the source of energy for the varieties of cold light by various prefixes. In chemiluminescence, for example, the source of energy is a chemical reaction.

Bioluminescence is a subcategory of chemiluminescence in which the energy-generating chemical reaction is enzyme catalyzed; enzymes are produced only by living systems.

Bioluminescence was described by early Chinese and Greek writers (e.g., Aristotle —De Anima), but little understanding of the process was achieved until late in the nineteenth century, although Robert Boyle had shown two centuries earlier that the process required oxygen, thereby implying a chemical reaction.³ In 1885, Dubois noted that a cold water extract of the luminous organ of a shellfish, although luminous initially, gradually darkened. Luminosity could be restored if a hot water extract of the same organ was added to the now inactive cold water extract.⁴ Dubois reasoned that an enzyme in the cold water extract consumed a substrate in the light-generating reaction. This substrate was present in the hot water extract, and adding it made further light generation possible until it too was consumed. He called the enzyme luciferase, and the substrate luciferin, from the Greek word (lucifer) for light-bearer. These are the names we have used ever since.

At the Paris International Exposition of 1900, Dubois exhibited a 24-1 jar of "Photobacteria" and nutrients that gave enough light for reading newspapers in an otherwise darkened room.⁵

One form of bioluminescence is the source of the light from the luminous beetles we call fireflies. Light generation in the firefly is stoichiometric with adenosine triphosphate (ATP) consumption with a quantum yield of 0.88.⁵ By contrast, the quantum yield from nonenzymic chemiluminescence reactions is seldom more than 0.01.⁶ Bacterial luciferase also produces light, but the chemistry of the reaction is quite different. In this case, light production depends upon the oxidation of reduced pyridine and flavin mononucleotides by oxygen in the presence of a long chain aldehyde.

Although both chemiluminescence and bacterial bioluminescence have important applications, particularly in clinical chemistry and in research, I will concentrate here on the ATP-dependent light generation from firefly luciferase, and on its applications to clinical microbiology. This topic has been reviewed in recent years by many well-qualified investigators whose efforts have greatly contributed to my own understanding of this fascinating field.^6-11

B. Mechanism Studies, Properties of Bioluminescence

It was more than a half century after Dubois that Professor William D. McElroy and his students at Johns Hopkins University began, in 1942, by repeating Dubois's experiments, then evolved a description of the chemistry of the firefly system that was nearly complete some 40 years later.¹²

Forty-seven years after he began his research in bioluminescence the redoubtable Professor McElroy, at this writing, remains active in highly original research.¹³

The reaction of light generation by the firefly system may be written:

luciferin + luciferase + ATP + Mg²⁺ →
luciferyladenylate-luciferase + pyrophosphate
lucifeiyladenylate-luciferase + O₂ →
luciferase + oxyluciferin + AMP + light + CO₂

ATP is nearly ...

Table of contents

• Cover
• Title
• Copyright
• Contents
• Chapter 1 Bioluminescence in Clinical Microbiology
• Chapter 2 Rapid Estimation of Microbial Numbers in Dairy Products Using ATP Technology
• Chapter 3 Rapid Food Microbiology: Application of Bioluminescence in the Dairy and Food Industry — A Review
• Chapter 4 The Preparation and Properties of Immobilized Firefly Luciferase for Use in the Detection of Microorganisms
• Chapter 5 Bioluminescence Applications in Brewing
• Chapter 6 Rapid Enumeration of Airborne Microorganisms by Bioluminescent ATP Assay
• Chapter 7 The Use of ATP Measurements in Biodeterioration Studies
• Chapter 8 Luminescence Instrumentation for Microbiology
• Index