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Wits SOP Flashcards

1
Q

Which bacteria commonly cause bacterial meningitis?

A

Aerobes

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2
Q

Which equipment and reagents are needed for CSF bench?

A

Microscope
Fuch’s Rosenthal counting chamber
Slides
Sterile glass Pasteur pipettes
Coverslips
India ink stain
CSF crystal violet
Glass tube
Capillary tubes
Centrifuge
Vortex
CO2 incubator

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3
Q

Must all CSFs be processed under the biosafety cabinet?

A

Yes

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4
Q

What do you do if an insufficient volume of CSF is received?

A

Priority
1. Inoculation of BHI
2. Blood agar
3. Chocolate agar
4. Gram
5. Cell count
6. Biochemistry

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5
Q

What does routine examination of CSF specimens include?

A
  1. Direct cell count
  2. Gram stain
  3. Biochemistry
  4. Culture

Other:
- India ink, CLAT, fungal culture
- TB, ADA
- serology
- viral studies
- bacterial antigen detection

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6
Q

What test can we do if neutrophils and lymphocytes on CSF specimen are difficult to differentiate?

A

Giemsa stain

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7
Q

Discuss the steps of CSF examination

A
  1. Record macroscopic CSF appearance
  2. Cell count
  3. Gram stain interpretation
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8
Q

How do you do a cell count using a Fuch’s Rosenthal counting chamber?

A
  1. Using a sterile Pasteur pipette deliver 9 drops CSF into clean tube
  2. Add 1 drop CSF CV to tube
  3. Mix well and fill counting chamber using a capillary tube
  4. Count all polymorphs, lymphocytes and erythrocytes (80 small squares = 5 large squares)
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9
Q

What do you do if the CSF is very bloody?

A

Dilute - deliver 8 drops of saline into a clean tube using a sterile pipette and using another sterile pipette add 1 drop of CSF and 1 drop of CSF crystal violet

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10
Q

How do you prepare the CSF for culture?

A
  1. Centrifuge remaining specimen for 10 minutes at 3000rpm
  2. Aspirate the supernatant with sterile pipette - send to chemistry
  3. Vortex sediment for at least 30 seconds to re-suspend pellet
  4. Inoculate media using sterile pipette with 1-2 drops of sediment and prepare 2 smears
  5. If organisms seen on gram stain and CSF remaining, perform Kirby-Bauer
  6. Air dry slides in biosafety cabinet
  7. Fix smears with heat/methanol
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11
Q

What temperature must the other CSF tubes be stored at in the freezer?

A

-15 to -25 degrees

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12
Q

How do you incubate the CSF cultures?

A

35-37 degrees in 5-10% CO2
Alternatives
- candle jar
- CO2 generating Gas-Pak

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13
Q

How do you process CSF culture?

A

72 hours re-incubating
MS and AST

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14
Q

What do you do if there is no growth on original culture plates but BHI is turbid?

A

Subculture onto blood and chocolate and perform a gram stain directly from broth

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15
Q

Which specimens must be sent on Dorsett egg slopes to NICD?

A

H. influenza
S. pneumo
N. meningitides

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16
Q

What do you expect concerning CSF macroscopically in acute bacterial infection?

17
Q

What do you expect concerning CSF macroscopically in TBM?

A

Clear
Spider clot formation
Slightly turbid

18
Q

What do you expect concerning CSF macroscopically in viral encephalitis?

19
Q

What do you expect concerning CSF macroscopically in neoplasm?

20
Q

What do you expect concerning CSF macroscopically in recent haemorrhage?

A

Bloody
Xanthochromic

21
Q

What do you expect concerning CSF macroscopically in old haemorrhage?

A

Xanthochromic
Hemolysed (pink)

22
Q

What do you expect concerning CSF macroscopically in cryptococcus?

23
Q

What do you expect concerning CSF cell count in acute bacterial infection?

A

Increased neutrophils
Variable lymphocytes

24
Q

What do you expect concerning CSF cell count in TBM?

A

Increased neutrophils
Increased lymphocytes

25
Q

What do you expect concerning CSF cell count in viral encephalitis?

A

Increased neutrophils
Normal lymphocytes

26
Q

What do you expect concerning CSF cell count in neoplasm?

A

Variable neutrophils
Variable lymphocytes

27
Q

What do you expect concerning CSF cell count in recent/old haemorrhage?

A

Variable neutrophils

28
Q

What do you expect concerning CSF cell count in cryptococcus?

A

Increase neutrophils
Increased lymphocytes

29
Q

What do you expect concerning CSF chemistry in acute bacterial infection?

A

Protein - high
Glucose - low
Chloride - normal

30
Q

What do you expect concerning CSF chemistry in TBM?

A

Protein - raised
Glucose - low
Chloride - low

30
Q

What do you expect concerning CSF chemistry in viral encephalitis?

A

Protein - slightly raised
Glucose - normal
Chloride - normal

31
Q

What do you expect concerning CSF chemistry in haemorrhage?

A

Protein - raised (blood cells)
Glucose - normal
Chloride - normal

32
Q

What do you expect concerning CSF chemistry in neoplasm?

A

Protein - raised
Glucose - normal
Chloride - normal

33
Q

What do you expect concerning CSF chemistry in cryptococcus?

A

Protein - raised
Glucose - low

CSF (1 decks)

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