biochemical id testing 2012 by miss sawsan flemban
TRANSCRIPT
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Biochemical ID Testing
Sawsan felmban
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Different kinds of bacteria are distinguished through….
Enzymes using biochemical tests
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Biochemical tests
Test the ability, activity or the presence of an
enzyme
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Biochemical ID Testing
• Catalase test.
• Coagulase test.
• Oxidase test.
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Catalase Test
• Reagents: H2O2
• Reaction :
• 2H2O2 2H2O + O2 (bubbles)
• Aim: to differentiate bacteria that produce catalase enzyme.
resu
lts
Bubbles +ve(staphylococcus)
No bubbles -ve
(NOT staph.)
Test QC:+ve cont.: Staphylococci spp.-ve cont.: Streptococci spp.
catalase
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Coagulase Test• Types of Coagulase:
• Free coagulase.
• Bound coagulase.
Reagent:
• Free coagulase: plasma containing coagulase reacting factor (CRF).
• Bound coagulase: plasma
• Principle:
fibrinogen fibrin clot
Aim: to ID S. aureaus
coagulase
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• Results:
• Free coagulase: fibrin clot (within 2hrs, 4hrs and 24 hrs) +ve
• Bound coagulase: fibrin clot 10 sec +ve.
• No fibrin clot: -ve
• Test QC:
• +ve Cont.: Staph. aureus
• -ve Cont.: Staph. epidermidis
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Free coagulase
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Bound coagulase
negative
posetive
posetive
negative
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Oxidase Test
• Reagent:
• 1% tetramethyl-p-phenylenediaminedihydrochloride.
• Principle:
• tetramethyl-p-phenylenediaminedihydrochloride + reduced cytochrome c
oxidized cytochrome c (Purple) + H2O
• Aim:
to ID cytochrome oxidase producing bacteria
oxidase
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Oxidase Test
resu
lts Dark purple : +ve
Pseudomonas
Colorless : - ve others