In same day sputum smear microscopy two sputum samples are collected with a gap of 1 hour. Study was conducted in department of Microbiology, GSL Medical College, Rajahmundry from March 2014 to May 2015. Three sputum (S, S2 and M) samples were collected from the pulmonary tuberculosis patients. Duplicate smears were prepared; one smear was stained by standard Ziehl Neelsen (ZN) staining technique and the other smear by reverse ZN (RZN) staining technique. In RZN staining technique, 0.1% methylene blue was used as primary staining reagent and 1% Carbol Fuchsin was the counter staining reagent. Of the 61 study participants, male/female ratio was 1.33:1, with mean age of 42.1 years. The smear positivity was 100%, 77% for spot morning (SM) approach and 92%, 72% for same day (SS2) approach, respectively for ZN staining and RZN staining. Statistically the difference was significant between the staining techniques in SM and SS2 approaches (P < 0.05). Due to the limited smear positivity, confusion in smear reading RZN staining technique neither used for teaching in tertiary hospitals nor implemented by the RNTCP.
Ziehl Neelsen (ZN) staining, Reverse Ziehl Neelsen (RZN) staining, Acid fast bacilli (AFB).
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