ACID-FAST STAINING

- Differentiate AFB from NAFB Hot Method Staining

- Mycobacterium spp. - Ziehl-Neelsen's Method
- Preliminary diagnosis of tuberculosis - Best acid-fast stain

- Monitor the effectiveness of treatments - Heat as the mordant

against tb (Isoniazid
- Procedure:

1. Flood the slide with Carbol Fuchsin solution,

Pre-analytical Phase using a low flame, and steam it for 5mins.

- Sterilization of working are using Lysol or
bleach to wipe the tabletops.
- KN95 Mask
2. Wash off excess stains with water.
3. Decolorize the smear by adding Acid Alcohol
in a small amount for 30 secs until no more
stain comes off.

4. Counterstain with methylene blue for 30secs.

Principle of AFS
5. Wash off excess stains with tap water.
- Mycolic acids in the cell walls give the afb cells
a higher affinity for the primary stain and 6. Read under the microscope.
resistance to decolorizer.
- Appearance:
- AFB- PINK; NAFB- BLUE
Colorless- Pink- Pink- Pink (AFB)

Colorless- Pink- Colorless- Blue (NAFB)

Stains
- Carbol fuchsin
Cold Method Staining
- Heat/Tergitol
- Kinyoun's Method
- Acid Alcohol
- Less sensitive than ZNM
- Methylene blue
- Does not use heat (Tergitol)

- Uses a slightly more lipid soluble and

Specimen collection and Smear concentrated carbol fuchsin.
preparation
Specimen: Sputum
Reporting (300 OIO fields)
- Early morning specimen 2-3 consecutive days
- CDC
- Placed in a sterile and enclosed container

- 2cmx1cm (=300 OIO fields)

- Coiling method
 Smear Smear Infectiousness
Result Interpretation to the Px
4+ Strongly Probably very
(>9/field) positive infectious
3+ (1- Strongly Probably very
9/field) positive infectious

2+ (1- Moderately Probably

9/10 positive infectious
fields)
1+ (1- Moderately Probably
9/100 positive infectious
fields)
+/-; (1- Weakly positive Probably
2/300 infectious
fields)
No AFB Negative Probably not
seen infectious*

-- - National Standard
Smear Report Smear Reading
0 No AFB seen
+N 1-9 AFB seen/100 fields

1+ 10-99 AFB seen/100 fields

2+ 1-10 AFB seen/50 fields
3+ More than 10 AFB
seen/field

Post-Analytical
- Sterilize all the materials and tools using lysol
or 10% bleach. All disposable materials
including ppe should be placed in their
designated trash bag.