Afb Sputum Staining

Sputum AFB – Procedure and Processing CRRI Skill Presentation Dr Irfan Ismail Ayub

Guide Dr Sandheep S MD, Assistant Professor

! ! " # $ %& '( +, -

"))*

!

Importance of AFB Sputum staining The DOTS program under RNTCP requires a good diagnostic tool for the success of the program. Sputum microscopy has 98 % case detection rate whereas Chest X-ray has 50 % detection rate only. Sputum AFB staining is also necessary for follow-up.

Give instructions – Rinse your mouth, stand facing a wall, away from wind, keep both hands on hip, cough forcibly, collect sputum in the mouth and spit carefully into the cup, close the lid. If no cup is available, any clean, wide mouthed container will do. Label the cup and lid with smear number. Improper labelling will lead to mix up of results. If mouth is not rinsed, or only saliva is given, it can lead to false positive / negative result.

Totally 3 sputum samples to be collected. Spot – morning – spot samples technique. For follow up – morning and spot samples.

. !

#

/ 0

. "12

( 0 !

. 3

! .

4

3

5 6

0

!

2 3

' !

7

$ ( 5

! .

'

8

9

2

! -

6 6

# !

/

+ -

3

$ / ) /

+ 4 1 ' "

/ 9 ,

!4

4 4 !

4 "

5. Microscopy Clean microscope and lens before use. Use plane mirror for focusing, raise condenser, open iris diaphragm fully and use oil immersion objective. Add a drop of cedar wood oil / liquid paraffin on smear. Focus using coarse adjustment till lens touches oil and then use fine adjustment. AFBs are seen as pink rods.

Microscopy - cont Follow a grid pattern to examine the smear. If grid pattern is not followed, whole smear will not be uniformly examined – bacilli may be missed. End

120

Start

Microscopy - cont Examine the smear for 5 min, covering about 100 fields. Examining for less than 5 min. can lead to missing of bacilli and false negative result.

6. Grading of Results Microscope

Result

Required fields

No bacilli seen in 100 fields 1-9 bacilli seen in 100 fields

Negative

100 fields

Actual no. of bacilli

100 fields

1+

100 fields

2+

50 fields

3+

10 fields

10-99 bacilli seen in 100 fields 100 or more in 100 fields 1-9 per field

Record results in register immediately. Clean the microscope lens after every positive smear and at end of work. Disposal of sputum cups and sticks by burning after a day’s work. Packing of smears – preserve all slides for supervision in a slide box. Positive and negative slides kept separately. Periodical reporting to be done – DTC to TO and SA everyday, and PHI to DTC on 5th of every month.

! &

' (

"#$ $$% )

*

Thank You