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Microbiologic Staining Method

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Title: Microbiologic Staining Method


1
Microbiologic Staining Method
  • ? ? ?
  • ? ? ? ?

2
Microbiological Staining
  • Gram stain
  • Acid fast stain
  • KOH stain
  • Wet mount
  • Indian ink

3
Gram Stain
  • Christian Gram, 1884
  • crystal violet
  • iodine-KI
  • alcohol wash
  • safranin

4
Significance
  • Culture
  • Buy time for golden period
  • Diagnosis

5
Interpretation
  • Morphological
  • Associate environment
  • Site-specific
  • Immunity-oriented

6
Morphologic interpretation
  • cocci vs bacilli, cocco-bacilli
  • Gram positive vs Gram negative

7
Morphologic interpretation
  • cocci vs bacilli, cocco-bacilli
  • Gram positive vs Gram negative

8
Gram-Positive Cocci
  • GPC in group
  • GPC in chain
  • GPC w/c capsule
  • GPC in pair
  • lancet shape
  • kidney shape
  • sarcinae

9
Gram-Positive Cocci
  • GPC in group
  • GPC in chain
  • GPC w/c capsule
  • GPC in pair
  • lancet shape
  • kidney shape
  • sarcinae

10
Gram Negative Cocci
  • Cocco-bacilli, Kidney-shape
  • G () vs G (-)

11
Bacilli
  • Gram positive bacilli
  • spore
  • spindle
  • box car
  • palisade
  • beaded
  • branching
  • Gram negative bacilli
  • pleomorphic
  • cocco-bacilli
  • capsule
  • curved

12
Bacilli
  • Gram positive bacilli
  • spore
  • spindle
  • box car
  • palisade
  • beaded
  • branching
  • Gram negative bacilli
  • pleomorphic
  • cocco-bacilli
  • capsule
  • curved

13
Presumptive Diagnosis
  • Staphylococcus aureus
  • Streptococcus pneumoniae
  • Hemophilus influenzae
  • Klebsiella pneumoniae
  • Nocardia spp
  • Candida spp

14
Associate environment
  • Neutrophils, epithelial cells
  • Aid for judgement of G(), G(-)
  • Intracellular vs extracellular
  • Mixed flora
  • w/o PMN, w/o bacteria
  • Non-bacteria organisms

15
Associate environment
  • Neutrophils, epithelial cells
  • Aid for judgement of G(), G(-)
  • Intracellular vs extracellular
  • Mixed flora
  • w/o PMN, w/o bacteria
  • Non-bacteria organisms

16
Associate environment
  • Neutrophils, epithelial cells
  • Aid for judgement of G(), G(-)
  • Intracellular vs extracellular
  • Mixed flora
  • w/o PMN, w/o bacteria
  • Non-bacteria organisms

17
Site specific
  • Gram negative cocci
  • urine
  • CSF
  • sputum
  • Gram positive bacilli
  • CSF
  • sputum
  • Gram positive cocci
  • urine

18
Site specific
  • Gram negative diplococci
  • urine N. gonorrheae
  • CSF N. meningitidis
  • sputum M. catarrhalis

19
Site specific
  • Gram positive bacilli
  • CSF L. monocytogenes
  • sputum Corynebaterium straitum
  • Gram positive cocci
  • urine Enterococcus , Viridans strep.

20
Immunity - oriented
  • Nosocomial infection
  • GNB in CSF
  • cocco-bacilli in sputum
  • T-cell immunity
  • GPB in CSF
  • Community-acquired
  • GNB in CSF
  • cocco-bacilli in sputum
  • T-cell immunity
  • GNB in joint

21
Immunity - oriented
  • Community-acquired
  • GNB in CSF
  • K. pneumoniae
  • cocco-bacilli sputum
  • H. influenzae
  • Nosocomial infection
  • GNB in CSF
  • P. aeruginosa
  • Enterobactericeae
  • cocco-bacilli in sputum
  • A. baumannii

22
Immunity - oriented
  • T-cell immunity
  • GPB in CSF
  • L. monocytogenes
  • T-cell immunity
  • GNB in joint
  • Salmonella spp.

23
Acid Fast Stain
  • Ziehl-Neelsen stain
  • Kinyoun modification
  • Mycobacteria
  • Nocardia, partial stain
  • NOT for actinomycosis
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