Comparison of real time IS6110-PCR, microscopy, and culture for diagnosis of tuberculous meningitis in a cohort of adult patients in Indonesia

PLoS One. 2012;7(12):e52001. doi: 10.1371/journal.pone.0052001. Epub 2012 Dec 21.

Abstract

Background: Bacteriological confirmation of tuberculous (TB) meningitis is difficult. Culture is slow and microscopy has insufficient sensitivity. We evaluated real time PCR targeting insertion sequence IS6110 among 230 consecutive adult patients with subacute meningitis in a referral hospital in Indonesia.

Methods: Cerebrospinal fluid (CSF) samples were examined using microscopy, solid and liquid culture, and real time IS6110-PCR with a fluorescence-labeled probe using DNA extracted from CSF. CSF samples from 40 non-infectious neurology patients were used as negative controls. IS6110-PCR results were linked with clinical and CSF characteristics.

Results: Most patients presented with subacute meningitis, after a median of 14 days of symptoms (range 7-30). After exclusion of cryptococcal and bacterial meningitis, 207 patients were classified as definite or probable TB meningitis; 17.9% with HIV infection. Among this group IS6110-PCR gave the highest positivity rate (68%, 95% CI 62-74%) compared with microscopy of ZN-stained slides (11%, 95% CI 7-15%), and mycobacterial culture using solid (36%, 95% CI 29-42%) and liquid (44%, 95% CI 37-51%) media. IS6110-PCR was positive in 92% of patients with culture-positive and 42% of patients with culture-negative probable TB meningitis. Among culture-negative patients, a positive PCR was associated with a history of TB treatment, a longer duration of illness, a higher CSF cell count and protein, and a lower CSF glucose. IS6110-PCR was negative in all CSF samples from non-meningitis control patients.

Conclusions: Real time IS6110-PCR is a quick, sensitive, and specific test for diagnosing of TB meningitis in this setting. Its performance in other (less-developed) settings needs further study.

Publication types

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

MeSH terms

  • Adult
  • Cohort Studies
  • Female
  • Humans
  • Indonesia
  • Male
  • Microscopy*
  • Mycobacterium tuberculosis* / cytology
  • Mycobacterium tuberculosis* / genetics
  • Mycobacterium tuberculosis* / growth & development
  • Real-Time Polymerase Chain Reaction*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tuberculosis, Meningeal / diagnosis*
  • Young Adult

Grants and funding

This study was financially supported by the Royal Academy of Arts and Sciences (KNAW, http://www.knaw.nl; 07-MP-10), The Netherlands and by IMPACT, a 5-year HIV program supported by the European Commission (SANTE/2005/105-033). Reinout van Crevel has a VIDI-grant from the Netherlands Organization for Scientific Research (NWO, http://www.nwo.nl; 017.106.310). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.