Study of respiratory symptoms among sputum positive

- Centrifuge.
- Vortex Mixer.
- Phenol can
- Electric slide warmer
• Procedure
A morning sputum sample or bronchial lavage was transferred to 50
ml plastic centrifuge tube, and an equal volume ofNALC-NaOH solution
was added and mixed well. The tube was allowed to stand for 15-18
minutes (not more than 20 minutes) at room temperature for
decontamination. Then, the sample was diluted to a 50 ml volume with a
sterile 0.067 M phosphate buffer saline (PBS), pH 6.8 to neutralize NaOH
and centrifuged at 2800 rpm at room temperature for 15 minutes. The
supernatant was poured off into a beaker containing 10% phenol, whereas
the deposit was resuspended in 2 ml of a sterile 0.2% bovine serum
albumin (BSA) in physiological saline (pH 6.8), then spread on the middle
2/3 of glass slide, after drying the slide on electric slide warmer at 65 C for
2 minutes (Ratnan and Marchy,1988).
c- Kinyoun '8 staining technique: It's modified Z. N (cold method) in
which:
1- high concentration ofcarbol fuchisin .
2- devoid of direct heat which has lethal effect ofTB bacilli,
3- The concentration of specimen by centrifugation conducted in this
method increase the capacity to detect mycobacteria (Saceanu et al.,
1993 and Fodor, 1995).
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