Application of fluorescent microscopy for improved detection of acid fast bacilli in sputum smear preparation

  • Dr. Archana Maruti Panage Assistant professor, Department of pathology, Rajiv Gandhi Medical College, Kalwa Thane, Maharashtra, India
  • Dr. Vivek D. Kharolkar Assistant professor, Grant Government Medical College, Mumbai, Maharashtra
  • Dr. Shreeram W. Chopade Associate professor, Department of pathology, Government Medical College, Akola, Maharashtra, India
Keywords: Tuberculosis, Ziehl-Neelsen, Auramine – O Staining, Fluorescence, Microscopy, Sputum smear.


Background: Tuberculosis remains a global health problem with an enormous burden of disease. Ziehl-Neelsen stain is the most commonly used method worldwide to detect tuberculous lymph node aspirates, however, Auramine – O Staining is being preferred owing to its high sensitivity, rapidity, and ease of screening. The aim of the present study was to compare the efficacy of fluorescent microscopy using Auramine O stain with conventional microscopy using Ziehl- Neelsen stain as a screening method to detect clinically suspected cases of pulmonary tuberculosis.

Method: A total of 100 clinically suspected cases of pulmonary tuberculosis referred to RNTCP laboratory from chest and TB OPD Medicine and Surgery and all other inpatient and out-patient departments were studied with detailed clinical history and sputum examination using Ziehl–Neelsen and Auramine O staining.

Results: Among 100 cases, 28% of cases were detected sputum smear-positive by Ziehl-Neelsen staining, and 30% cases were detected by Auramine O stain using a fluorescence microscope. Maximum sputum smears positive (36.6%) cases were in the age group of 31-40 years with male predominance (75%). Auramine O stain showed higher grading in 4 cases in which one case was scanty on Ziehl-Neelsen smear showed positive 1+ by Auramine O and other 2 cases which were negative by Ziehl-Neelsen stain were showed 1+ result on Auramine O stain. One case which showed 2+ grading on Ziehl-Neelsen stain showed 3+ grading with Auramine O stain. Fluorescence microscopy using Auramine O showed better grading than the Ziehl-Neelsen stain.

Conclusion: Fluorescence microscopy is more superior in detecting sputum smear-positive cases over Ziehl-Neelsen stain.


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DOI: 10.26838/MEDRECH.2021.8.5.559
Published: 2021-10-21
How to Cite
Panage AM, Kharolkar VD, Chopade SW. Application of fluorescent microscopy for improved detection of acid fast bacilli in sputum smear preparation. Med. res. chronicles [Internet]. 2021Oct.21 [cited 2022May22];8(5):439-47. Available from:
Original Research Article