Selvakumar, NRavikumar, DSivagamasundari, SGopi, P GNarayanan, P R2006-06-042009-05-272006-06-042009-05-272006-06-04Selvakumar N, Ravikumar D, Sivagamasundari S, Gopi PG, Narayanan PR. A novel method of staining acid-fast bacilli in sputum containers. Indian Journal of Medical Research. 2006 Jun; 123(6): 776-80http://imsear.searo.who.int/handle/123456789/22782BACKGROUND & OBJECTIVES: Making centrifuged deposit smears from sputum to detect acid-fast bacilli (AFB) is considered hazardous. We carried out this study to stain the centrifuged deposits with carbol-fuchsin in sputum containers and to decolourize and counterstain their smears made on glass slides. METHODS: The centrifuged deposits of 180 sputum samples from pulmonary tuberculosis patients were used for making smears (initial deposit smears) and staining by Ziehl-Neelsen (ZN) method for the detection of AFB. Each of the sputum deposit was then treated with one ml of 1 per cent carbol-fuchsin and a smear made between 2 to 3 h was then decolourized and counterstained by the same procedures followed in ZN method (2 h stained deposit smear). The coded initial deposit smears and the corresponding 2 h stained deposit smears were read by the same readers and the results compared. RESULTS: One hundred and fifty (70 positive and 80 negative) 2 h stained deposit smears were compared with initial deposit smears and the difference was not statistically significant. INTERPRETATION & CONCLUSION: Centrifuged deposits of sputum in sputum containers can be stained by carbol-fuchsin within 2-3 h and their smears made subsequently on glass slides can then be decolourized and counterstained by the procedures followed in ZN method for detection of AFB by light microscopy.engCentrifugationHumansMicroscopyMycobacterium --isolation & purificationRosaniline DyesSpecimen Handling --methodsSputum --microbiologyStaining and Labeling --methodsTuberculosis, Pulmonary --diagnosisJournal Article