An operational study comparing microscopes and staining variations for tuberculosis LED FM

A Van Deun, K J M Aung, M H Khan, Bouke C de Jong, M Gumusboga, M A Hossain

Research output: Contribution to journalA1: Web of Science-articlepeer-review


SETTING: Tuberculosis control projects, Damien Foundation Bangladesh.

OBJECTIVES: To compare transmitted fluorescence (Olympus CX21™/FRAEN FluoLED™) with epi-fluorescence (Zeiss Primostar iLED™) light-emitting diode microscopes (LED-FM) and various auramine staining and destaining/counterstaining techniques for the detection of acid-fast bacilli.

DESIGN: Multicentre blinded reading of routine smears on both types of microscopes using different staining techniques in multiple phases. LED-FM rechecking of discordant series with and without restaining to calculate operating characteristics.

RESULTS: Among 64 874 smears, both instruments detected 9.6% positives. Compared to the standard technique, the stronger auramine-O formulation did not perform better. Thiazine red counterstaining tended to yield more false-positive as well as false-negative errors. Combined destaining/counterstaining (sensitivity 93%, positive predictive value [PPV] 98%) proved significantly less effective. Both destaining with 1% hydrochloric acid (HCl) and 10% alcohol and the standard 0.5% HCl and 70-95% alcohol were equally accurate (sensitivity 95-96%, PPV 99%). The sturdiness of the microscopes in field conditions was sub-optimal: only 5/16 instruments did not break down.

CONCLUSIONS: Both microscopes performed equally well. The standard staining technique is as good as the more complicated and expensive variations. A destaining solution containing only 10% alcohol works perfectly well. The inferior quality of LED-FM microscope components is an obstacle to FM expansion.

Original languageEnglish
JournalInternational Journal of Tuberculosis and Lung Disease
Issue number8
Pages (from-to)964-971
Number of pages8
Publication statusPublished - 2014


  • Benzophenoneidum
  • False Negative Reactions
  • False Positive Reactions
  • Microscopy, Fluorescence
  • Organic Chemicals
  • Predictive Value of Tests
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sputum
  • Staining and Labeling
  • Tuberculosis


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