Optimising Mycobacterium tuberculosis detection in resource limited settings

BMJ Open. 2014 Mar 3;4(3):e004093. doi: 10.1136/bmjopen-2013-004093.

Abstract

Objectives: The light-emitting diode (LED) fluorescence microscopy has made acid-fast bacilli (AFB) detection faster and efficient although its optimal performance in resource-limited settings is still being studied. We assessed the optimal performances of light and fluorescence microscopy in routine conditions of a resource-limited setting and evaluated the digestion time for sputum samples for maximum yield of positive cultures.

Design: Cross-sectional study.

Setting: Facility-based involving samples of routine patients receiving tuberculosis treatment and care from the main tuberculosis case referral centre in northern Nigeria.

Participants: The study included 450 sputum samples from 150 new patients with clinical diagnosis of pulmonary tuberculosis.

Methods: The 450 samples were pooled into 150 specimens, examined independently with mercury vapour lamp (FM), LED CysCope (CY) and Primo Star iLED (PiLED) fluorescence microscopies, and with the Ziehl-Neelsen (ZN) microscopy to assess the performance of each technique compared with liquid culture. The cultured specimens were decontaminated with BD Mycoprep (4% NaOH-1% NLAC and 2.9% sodium citrate) for 10, 15 and 20 min before incubation in Mycobacterium growth incubator tube (MGIT) system and growth examined for acid-fast bacilli (AFB).

Results: Of the 150 specimens examined by direct microscopy: 44 (29%), 60 (40%), 49 (33%) and 64 (43%) were AFB positive by ZN, FM, CY and iLED microscopy, respectively. Digestion of sputum samples for 10, 15 and 20 min yielded mycobacterial growth in 72 (48%), 81 (54%) and 68 (45%) of the digested samples, respectively, after incubation in the MGIT system.

Conclusions: In routine laboratory conditions of a resource-limited setting, our study has demonstrated the superiority of fluorescence microscopy over the conventional ZN technique. Digestion of sputum samples for 15 min yielded more positive cultures.

Keywords: Microbiology.

Publication types

  • Evaluation Study

MeSH terms

  • Cross-Sectional Studies
  • Health Resources*
  • Humans
  • Lung / microbiology*
  • Microscopy, Fluorescence / methods*
  • Mycobacterium tuberculosis / growth & development*
  • Nigeria
  • Sensitivity and Specificity
  • Sputum / microbiology*
  • Tuberculosis, Pulmonary / diagnosis*
  • Tuberculosis, Pulmonary / microbiology