Abstract

BackgroundDirect smear microscopy using Ziehl-Neelsen (ZN) staining is the mainstay of tuberculosis (TB) diagnosis in most high burden countries, but is limited by low sensitivity in routine practice, particularly in high human immunodeficiency virus (HIV) prevalence settings.MethodsWe compared the performance of three commercial light emitting diode (LED)-based microscopy systems (Primostar™ iLED, Lumin™ and AFTER®) for fluorescent detection of Mycobacterium tuberculosis with ZN microscopy on slides prepared from sputum of TB suspects. Examination time for LED-based fluorescent microscopy (LED FM) and ZN slides was also compared, and a qualitative user appraisal of the LED FM systems was carried out.ResultsLED FM was between 5.6 and 9.4% more sensitive than ZN microscopy, although the difference was not statistically significant. There was no significant difference in the sensitivity or specificity of the three LED FM systems, although the specificity of Fraen AFTER was somewhat lower than the other LED FM methods. Examination time for LED FM was 2 and 4 times less than for ZN microscopy. LED FM was highly acceptable to Ugandan technologists, although differences in operational performance of the three systems were reported.ConclusionsLED FM compares favourably with ZN microscopy, with equivalent specificity and a modest increase in sensitivity. Screening of slides was substantially quicker using LED FM than ZN, and LED FM was rated highly by laboratory technologists. Available commercial systems have different operational characteristics which should be considered prior to programmatic implementation.

Highlights

  • Direct smear microscopy using Ziehl-Neelsen staining is still the mainstay of tuberculosis (TB) diagnosis in most high burden countries, including Uganda, having remained essentially unchanged for over 100 years

  • In total, 1 low false positive (LFP) was obtained for ZN, 0 errors were obtained for iLED, 4 LFPs and 1 low false negative (LFN) were obtained for Fraen AFTER and 1 LFP was obtained for Lumin

  • Sensitivity of the light emitting diode (LED) fluorescent microscopy (FM) methods was between 5.6% and 9.4% higher than ZN

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Summary

Introduction

Direct (un-concentrated) smear microscopy using Ziehl-Neelsen staining is still the mainstay of tuberculosis (TB) diagnosis in most high burden countries, including Uganda, having remained essentially unchanged for over 100 years. This method is rapid and inexpensive and highly specific for Mycobacterium tuberculosis in high burden settings. The main limitation of the method is its low sensitivity in programmatic settings, in human immunodeficiency virus (HIV) co-infected patients [1]. Direct smear microscopy using Ziehl-Neelsen (ZN) staining is the mainstay of tuberculosis (TB) diagnosis in most high burden countries, but is limited by low sensitivity in routine practice, in high human immunodeficiency virus (HIV) prevalence settings

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