ABSTRACT
Introduction .................................................................................................................................... 136 Culture ............................................................................................................................................ 136
Media Types .............................................................................................................................. 136 General Media ...................................................................................................................... 136 Selective Media .................................................................................................................... 136 Differential Media ................................................................................................................ 136
Colonial Morphology ................................................................................................................ 137 Gram Stain ................................................................................................................................ 137 Biochemical Tests ..................................................................................................................... 137
Immunology-Based Assays ............................................................................................................ 139 Agglutination Assays ................................................................................................................ 139 Antibody Labeling Techniques ................................................................................................. 139 Enzyme Immunoassay Generations .......................................................................................... 140 Antibody Technique Variations ................................................................................................. 141
Second Antibody .................................................................................................................. 141 Enhanced EIA....................................................................................................................... 142 Biotin Avidin ........................................................................................................................ 142 Magnetic Beads .................................................................................................................... 142 Neutralization ....................................................................................................................... 142 Slide Tests ............................................................................................................................. 142
Nucleic Acid Probes ....................................................................................................................... 143 Sample Amplication Methods...................................................................................................... 143
Polymerase Chain Reaction ...................................................................................................... 143 Real-Time PCR ......................................................................................................................... 143 Taqman ...................................................................................................................................... 144 Light Cycler .............................................................................................................................. 144 Variations of PCR ...................................................................................................................... 145 Ligase Chain Reaction .............................................................................................................. 145 Other Sample Amplication Methods ....................................................................................... 145
Signal Amplication Methods ....................................................................................................... 146 Branched-Chain DNA ............................................................................................................... 146 Hybrid Capture .......................................................................................................................... 147
DNA Microarray Technology ........................................................................................................ 147 Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) ........................................................................................................................ 147 Electrophoresis ............................................................................................................................... 148 References ...................................................................................................................................... 149
Microbiologists have been identifying organisms, since Pasteur rst realized that in addition to producing the ne wines of France, microorganisms also caused disease. Since that time, it appears as if the chief goal of diagnostic microbiology has been to come up with the one test that would separate the helpful organisms from the harmful ones. The unfortunate thing is that it seems that every time someone developed this ultimate test, eventually it was discovered that there was always some overlap. The purpose of this chapter is to describe the major methods being used to identify microorganisms in the laboratory today.
