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Bronchoalveolar Lavage Enzyme-linked Immunospot for a Rapid Diagnosis of Tuberculosis

A Tuberculosis Network European Trialsgroup Study

¹ Clinical Infectious Diseases, Research Center Borstel, Borstel Germany; ² Medical Microbiology, Diakonessenhuis, Utrecht, The Netherlands; ³ Hygiene and Preventive Medicine Institute, University Sassari, Italy; ⁴ Translational Research Unit, National Institute for Infectious Diseases, Rome, Italy; ⁵ Department of Microbiology, Hospital Universitari Germans Trias i Pujol, Ciber Enfermedades Respiratorias, Institutio de Salud Carlos III, Badalona, Spain; ⁶ Department of Oncology, Hematology and Respiratory Diseases, University of Modena and Reggio Emilia, Modena, Italy; ⁷ Pulmonary and Critical Care Medicine, University Heidelberg Thoraxclinic, Heidelberg, Germany; ⁸ Pneumology, Hospital Großhansdorf, Großhandorf, Germany; ⁹ Pulmonary Medicine, Diakonessenhuis, Utrecht, The Netherlands; ¹⁰ WHO Collaborating Centre for TB and Lung Diseases, Fondazione S. Maugeri, Care and Research Institute, Tradate, Italy; ¹¹ Immune Cell-Analytics, Research Center Borstel, Borstel, Germany

Correspondence and requests for reprints should be addressed to Christoph Lange, M.D., Ph.D., Research Center Borstel, Parkallee 35, D-23845 Borstel, Germany. E-mail: clange{at}fz-borstel.de

Rationale: The rapid diagnosis of pulmonary tuberculosis (TB) is difficult when acid fast bacilli (AFB) cannot be detected in sputum smears.

Objectives: Following a proof of principle study, we examined in routine clinical practice whether individuals with sputum AFB smear-negative TB can be discriminated from those with latent TB infection by local immunodiagnosis with a Mycobacterium tuberculosis–specific enzyme-linked immunospot (ELISpot) assay.

Methods: Subjects suspected of having active TB who were unable to produce sputum or with AFB-negative sputum smears were prospectively enrolled at Tuberculosis Network European Trialsgroup centers in Europe. ELISpot with early-secretory-antigenic-target–6 and culture-filtrate-protein–10 peptides was performed on peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage mononuclear cells (BALMCs). M. tuberculosis–specific nucleic acid amplification (NAAT) was performed on bronchoalveolar lavage fluid.

Measurements and Main Results: Seventy-one of 347 (20.4%) patients had active TB. Out of 276 patients who had an alternative diagnosis, 127 (46.0%) were considered to be latently infected with M. tuberculosis by a positive PBMC ELISpot result. The sensitivity and specificity of BALMC ELISpot for the diagnosis of active pulmonary TB were 91 and 80%, respectively. The BALMC ELISpot (diagnostic odds ratio [OR], 40.4) was superior to PBMC ELISpot (OR, 10.0), tuberculin skin test (OR, 7.8), and M. tuberculosis specific NAAT (OR, 12.4) to diagnose sputum AFB smear-negative TB. In contrast to PBMC ELISpot and tuberculin skin test, the BALMC ELISpot was not influenced by previous history of TB.

Conclusions: Bronchoalveolar lavage ELISpot is an important advancement to rapidly distinguish sputum AFB smear-negative TB from latent TB infection in routine clinical practice.

AT A GLANCE COMMENTARY Scientific Knowledge on the Subject The rapid diagnosis of active pulmonary tuberculosis is difficult when acid-fast bacilli (AFB) cannot be detected in sputum smears. Local immunodiagnosis by Mycobacterium tuberculosis–specific enzyme-linked immunospot assay (ELISpot) is a promising method for the rapid identification of patients with sputum AFB smear-negative tuberculosis. What This Study Adds to the Field In a prospective multicenter TBNET-study, patients with sputum AFB smear-negative pulmonary tuberculosis could rapidly be distinguished from patients with latent tuberculosis infection by the M. tuberculosis–specific ELISpot on cells from the BAL fluid with a high diagnostic sensitivity and specificity. These findings may have significant implications for the rapid decision to initiate antituberculosis treatment where bronchoscopy is routinely performed for individuals suspected to be affected by sputum AFB smear-negative tuberculosis.

 

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