title: Direct detection of Mycobacterium tuberculosis in sputum: A validation study using solid phase extraction-gas chromatography-mass spectrometry.
abstract: Tuberculosis (TB) remains a worldwide health problem, especially in developing countries. Correct identification of Mycobacterium tuberculosis (MTB) infection is extremely important for providing appropriate treatment and care to patients. Here we describe a solid phase extraction-gas chromatography-mass spectrometry method (SPE-THM-GC-MS) for the detection of five biomarkers for M. tuberculosis. The method for classification is developed and validated through the analysis of 112 sputum samples from patients suspected of having TB. Twenty of twenty-five MTB culture-positive sputum samples were correctly classified as positive by our improved SPE-THM-GC-MS method. Eighty-five of eighty-seven MTB culture-negative samples were also negative by SPE-THM-GC-MS. The overall sensitivity of the new SPE-THM-GC-MS method is 80% (20/25) and the specificity is 98% (85/87) compared with culture. The method proved to be reliable and, although complex in principle, easy to operate due to the high degree of automation.
authors.affiliation: University of Amsterdam, Van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, P.O. Box 94157, 1090 GD Amsterdam, The Netherlands. Electronic address: M.PachecoBotelhoMourao@uva.nl.
authors.forename: Marta P B
authors.initials: MP
authors.lastname: Mourão
authors.affiliation: University of Amsterdam, Van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, P.O. Box 94157, 1090 GD Amsterdam, The Netherlands.
authors.forename: Sjoukje
authors.initials: S
authors.lastname: Kuijper
authors.affiliation: University of Amsterdam, Van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, P.O. Box 94157, 1090 GD Amsterdam, The Netherlands.
authors.forename: Ngoc A
authors.initials: NA
authors.lastname: Dang
authors.affiliation: Desmond Tutu TB Centre, Stellenbosch University, P.O. Box 19063, Tygerberg 7505, South Africa.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Walters
authors.affiliation: University of Amsterdam, Van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, P.O. Box 94157, 1090 GD Amsterdam, The Netherlands; Unilever Research and Development, P.O. Box 114, 3130 AC Vlaardingen, The Netherlands.
authors.forename: Hans-Gerd
authors.initials: HG
authors.lastname: Janssen
authors.affiliation: University of Amsterdam, Van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, P.O. Box 94157, 1090 GD Amsterdam, The Netherlands.
authors.forename: Arend H J
authors.initials: AH
authors.lastname: Kolk
mesh_terms: D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*; D013183:Sputum / Q000382:microbiology*; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology
keywords: Direct detection; Gas chromatography–mass spectrometry; Mycobacterium tuberculosis; Solid Phase Extraction; Validation