Auramine phenol staining of smears for screening acid fast bacilli in clinical specimens.

Tuberculosis is a major threat to the public health allover world. Out of the total tuberculosis cases reported globally, more than half are reported from African continent and India. Two to three fold rises in tuberculosis cases has been reported in the last two decades. Early diagnosis and treatment is one of the effective tools to control the rapid spread of disease. The aim of this study was to find out the value of auramine phenol (AP) staining technique in diagnosis of the suspected tuberculosis cases. A total of 2000 samples which included sputum (746), gastric aspirates (380), urine (336), endometrial biopsy (150), pleural fulids (146), Synovial fluids (67), ascitic fluids (35), cerebrospinal fluids (43), bone marrow (18), lymph node biopsy (11), pericardial aspirates (6), skin biopsy (4), peritoneal fluids (2), and stool (1) were included in the study. Sample were subjected for decontamination procedure by using standard Petroffs method. The deposit smears were stained by auramine phenol (AP stain) and Ziehl-Neelsenstaining (ZN stain) and specimens were cultured for Mycobacterium tuberculosis. Of the total positive isolates 69.23% were having pulmonary tuberculosis and 30.76 had extrapulmonary tuberculosis Genitourinary tuberculosis was the most common diagnosis among the extrapulmonary tuberculosis followed by chronic synovitis, bursitis, meningitis, septic arthritis and pericardial effusion. Out of 130 positive samples 70 by culture, 66 smears were positive by auramine phenol stain and 62 were positive by ZN stain. A total of 27 samples were tested positive only by AP staining technique, which included (12) pulmonary and (15) extrpulmonary samples. The endometrial biopsy and pericardial fluid samples showed positive for acid fast bacilli by AP stain only, whereas ZN stain and culture technique failed to demonstrate any bacilli in the same sample. Auramine stain showed high sensitivity (47.14%) and specificity (96.58%). Result of the present study showed that the auramine stain is a better method for screening samples from the suspected cases of tuberculosis sample especially pulmonary and extrapulmonary cases where bacilli count is usually low.

The value of fluorescence microscopy of auramine stained sputum smears for the diagnosis of pulmonary tuberculosis.

Laboratory diagnosis of pulmonary tuberculosis rests on the bacteriological examination of sputum smears stained by the Ziehl-Neelsen (ZN) method for acid fast bacilli (AFB). In the present study, we have compared light microscopy of ZN stained smears with that of fluorescence microscopy of sputum smears stained by auramine-phenol flurochrome dye for detection of AFB in sputum specimens. Sputum specimens from a total of 2,600 clinically suspected and diagnosed cases of pulmonary tuberculosis were examined by both the methods. Sputum specimens from a total of 1,104 patients were found to be positive for AFB. These included sputa from 975 (37.5%) patients positive for AFB by both ZN and auramine staining methods and sputa from an additional 129 (4.96%) patients positive for AFB by auramine staining only. Thus auramine staining of sputum smears in comparison to that of ZN staining is a better method of sputum microscopy for demonstration of AFB in sputum specimens. Fluorescence microscopy is relatively more sensitive and has the added advantage of allowing a large number of sputum specimens to be examined in a given time, in laboratories equipped with a fluorescent microscope.

Auramine staining in histopathology sections.

Auramine staining was done on 65 histopathological sections from different types of treated leprosy cases which were negative by Fite-Farraco stain. All the sections except one showed auramine positive organisms. The organisms were mostly coccoid except in BL/LL cases where beaded bacilli could be seen.