Conventional and molecular techniques in the diagnosis of pulmonary tuberculosis: a comparative study

J Bras Pneumol. 2008 Dec;34(12):1056-62.
[Article in English, Portuguese]

Abstract

Objective: To compare four laboratory methods in the diagnosis of pulmonary tuberculosis.

Methods: Respiratory secretion specimens were collected from 160 patients suspected of having pulmonary tuberculosis. Direct testing for Mycobacterium tuberculosis was carried out using Ziehl-Neelsen and auramine staining. In addition, culture in Löwenstein-Jensen (LJ) medium and polymerase chain reaction (PCR) were used. The strains isolated were identified by means of a radiometric method using p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) and classical methods. The sensitivity of the methods was compared to the gold standard for the diagnosis of pulmonary tuberculosis, based on clinical, radiological and microbiological criteria.

Results: Of the 160 patients, 142 were diagnosed with pulmonary tuberculosis according to the gold standard. The sensitivity of Ziehl-Neelsen staining, auramine staining, culture in LJ medium and PCR was 54.2%, 58.4%, 67.6% and 77.5%, respectively, when compared with the diagnostic criterion adopted. All four methods presented 100% specificity. In the identification of mycobacteria, there was high (96.8%) concordance between PCR and the radiometric method using NAP. The sensitivity of PCR was 50.8% in samples with negative sputum smear microscopy results and 98.8% in those with positive results. The sensitivity of PCR was lower in specimens with negative results in sputum smear microscopy and culture than in those with positive results (25.6% and 99.0%, respectively).

Conclusions: We found PCR to be a promising method for the diagnosis of pulmonary tuberculosis, even in paucibacillary specimens. Simultaneous identification and faster results are additional advantages of this method.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Bacteriological Techniques / methods*
  • Benzophenoneidum
  • Coloring Agents
  • Culture Media
  • Data Interpretation, Statistical
  • Female
  • Humans
  • Hydroxypropiophenone / analogs & derivatives
  • Male
  • Middle Aged
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Sputum / microbiology*
  • Staining and Labeling / methods*
  • Tuberculosis, Pulmonary / diagnosis*
  • Tuberculosis, Pulmonary / genetics
  • Tuberculosis, Pulmonary / microbiology
  • Young Adult

Substances

  • Coloring Agents
  • Culture Media
  • Hydroxypropiophenone
  • Benzophenoneidum
  • 4-nitro-alpha-acetylamino-beta-hydroxypropiophenone