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Published ahead of print on July 20, 2006, doi:10.1164/rccm.200604-465OC

Am. J. Respir. Crit. Care Med., Volume 174, Number 9, November 2006, 1048-1054

A more recent version of this article appeared on November 1, 2006
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Submitted on April 3, 2006
Accepted on July 19, 2006

Rapid Diagnosis of Smear-negative Tuberculosis by Bronchoalveolar Lavage Enzyme-linked Immunospot

Claudia Jafari1, Martin Ernst2, Roland Diel3, Ulf Greinert1, Barbara Scheuerer1, Detlef Kirsten4, Kathleen Marienfeld2, Ajit Lalvani5, and Christoph Lange1*

1 Division of Clinical Infectious Diseases, University of Dusseldorf, Borstel, Germany, 2 Division of Immune Cell Analytics, University of Dusseldorf, Research Center Borstel, Borstel, Germany, 3 Department of Public Health, University of Dusseldorf, Dusseldorf, Germany, 4 Center for Pulmonary Medicine and Thoracic Surgery, Hospital Grosshansdorf, Grosshansdorf, Germany, 5 Nuffield Department of Clinical Medicine, Tuberculosis Immunology Group, University of Oxford, John Radcliff Hospital, Oxford, United Kingdom

* To whom correspondence should be addressed. E-mail: clange{at}fz-borstel.de.

Rationale: A large proportion of patients with active pulmonary tuberculosis (pTB) has negative acid-fast bacilli (AFB) smear results for sputum and bronchial secretions (BS). Detectable growth of M. tuberculosis (MTB) in cultures takes several weeks and MTB-specific DNA-amplification results on sputum and BS are variable in these patients. Objectives: We investigated whether a rapid diagnosis of pTB can be established by enumeration of MTB-specific mononuclear cells from the bronchoalveolar lavage (BAL) in routine clinical practice. Methods: Patients presenting to a tertiary hospital with medical histories and pulmonary infiltrates compatible with tuberculosis, and negative AFB smear results (3) from the sputum were prospectively enrolled in this study. A MTB-specific ELISPOT (T-SPOT-TB) with ESAT-6 and CFP-10 peptides was performed on peripheral blood mononuclear cells (PBMCs) and mononuclear cells from the BAL (BALMCs). Measurements and Main Results: Out of 37 patients, 12 were found to have smear-negative pTB and 25 were found to have an alternative diagnosis. Patients with tuberculosis had a median number of 17 ESAT-6 and 24.5 CFP-10 specific cells/200000 PBMCs and 37.5 ESAT-6 and 49.5 CFP-10 specific cells/200000 cells in the BAL. Control patients had a median of 1 ESAT-6 and 1 CFP-10 specific cells/200 000 PBMCs and 0 ESAT-6 and 0 CFP-10 specific cell/200 000 cells in the BAL (p<0,0001). All patients with TB but none of the controls had more than 5 spot-forming cells/200000 with either peptides in the BAL ELISPOT. Conclusions: Smear-negative pulmonary tuberculosis can rapidly be diagnosed by identification of MTB-specific cells in the BAL.
Key words: tuberculosis, BAL, ELISPOT, ESAT-6, CFP-10, T-SPOT-TB




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