----- Active space BIOCHEMICALS AND ANTIBIOTIC SENSITTVIT
TESTS
Biochemical Reactions [Link]
Catalase test: Oxidase test:
" To detect catalase enzume. " To detect cutochrome c oxidose in bacteria.
catalase
" Bacteriol suspension + HO HO+O, Filter paper strip impregnated uwith Kovac's
"Catalase tve :most pathogenic bacteria. reagent.
Except: " Oxidase tve:All patthoqenic bacteria.
[Link]. Except:
- Streptococcaceae. - Corynebacterium. -Staphyjococoi.
- Enterobacteraceae.
-Shigela dysentery type . - Streptococci.
Positive Négative
Oxidase test
Catalase test +ve
qlucose utilization test:
Detect glucose utilization.
medium : Hugh Leifson oxidative-fermentative medium
(contains glucose).
Principle:
Glucose utilization Produced acid: Tuns yellow.
-Aerobic
" Available as pair of test tubes
Anaerobic.
Inference:
"color change:
D
Fedback - In only aerobic test tube : Oxidative utilization of glucose.
- In both aerobic and anaerobic test tubes : Fermentative utlization of qlucose.
somplesi
-Focultotive anaerobes iFerment sugars.
eg ienterobacter iaceae, Staphylococci, Streptococci.
Microblology v3,0 Marrow 8.0 2024 Page /4
-Strict aerobes : Oxidise sugars. Active space ----
&.q :Pseudomonas, rucella, Bordetella, Nocardia.
F F O F
-Oxidative utilisation
of glucose
Fermentative
utlisation of glucose
[Link] @[Link]
Glucose utilisation
Urease test: Imvic test:
Procedure: Indole test:
Bacteria
Bacteria produce urease. "Tryptophan Indole.
urea urease NH, t CO,
(Produced by Indole Test Methyl Red Test VP Test
bacteria) Citrate Agar
NH
Alkaline medium
Urease test
medium:Christensen's medium.
urease positive organisms, :
D
Fedback
mnemonic : PUNCH MSKe
-Proteus.
-ureaplasma.
- Nocordia
-morganelloa.
-S. oureus, S. saprophyticus.
-Klebsiela.
-cryptocoeeus -erucella.
-H. pylor
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Active space Antibiotic Sensitivity Testing 00:1
medium:
Non-fastidious bacteria:Cation adjusted mueler Hinton agar/broth.
Fastidious bacteria :Cation adjusted mueller Hinton agar/broth +5% blood.
Anaerobic bacteria : Wilkins chalaren agar. O.5 mcF ImcF IS mcF
Standard inoculum of test bacteria !
Sample turbidity compared with standard tubes.
Turbidity is [Link]- Antibiotic sensitivity testing can be done.
Other parameters :
[Link]@[Link] Temperature : 35-37°C.
Except: S. aureus (30-33°c) as meSA is
detected best at louw temperatures. mcFarland standoard tubes
Time of interpretation : Io-18 hours.
meTHODS OF AST
Phenotypic : manual
moleculor:
" Dilution
Automated Detecting genes.
Disc difusion.
" epsilometer method.
" chrom agar.
Dilution method o AST: Agar dilution
macro broth dilution
1. Broth dilution a Aqar dilution.
micro broth dilution
Procedure:
"mic ninimuminhibitory Concentratiorn) :Lowest concentration o
antibiotic uwhich just inhibits grouth of bacterium. 37 °c miero dilution
Doubling dilutions of antibodies + standard inoculum mIC.
Resistant
Compare with break point concentration
-Sensitive
Advantoge:Determines mic (Reerence method).
D
Fedback
Disadvantoge: Loborious.
Disc diffusion method:
Kirby Bouer (m/) Stokes
ProcedureiDisc impregnated uwith single
stondordised concentration of antibiotic.
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Active space
Test Control
BEFORE GROWTH AFTER GROWTH
Growth Time
24 hous
No bacterial growth
Antibiotic disks
Agarmedia, spread (Zone of inhibition)
on Petri dish
Bacterial growth
herby Bauer disc diffusion metthod Stokes disc difSusion method
[Link] i Easy
Disadvantage : Cannot determine exact mic.
epsilometer/e test: Chromogenic method:
" Dilution + disc diffusion method. Detects:
" Reference: Dilution method. " mRSA.
" eseL producing gram negative bacteria.
mIc
Epsilometer/e meter
Automated methods:
Based on micro broth dilution method.
L. Viteka.
a. Phoenik method.
Chromogenic method
molecular methods:
Detects oenes.
meSA mecA gene. [Link] resistanceirpoe gene.
DGe by'
enplytyng method. " Nucleic acid seqencing
Hyrdisation probes.
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