title: Volatile biomarkers of pulmonary tuberculosis in the breath.
abstract: Pulmonary tuberculosis may alter volatile organic compounds (VOCs) in breath because Mycobacteria and oxidative stress resulting from Mycobacterial infection both generate distinctive VOCs. The objective of this study was to determine if breath VOCs contain biomarkers of active pulmonary tuberculosis. Head space VOCs from cultured Mycobacterium tuberculosis were captured on sorbent traps and assayed by gas chromatography/mass spectroscopy (GC/MS). One hundred and thirty different VOCs were consistently detected. The most abundant were naphthalene, 1-methyl-, 3-heptanone, methylcyclododecane, heptane, 2,2,4,6,6-pentamethyl-, benzene, 1-methyl-4-(1-methylethyl)-, and cyclohexane, 1,4-dimethyl-. Breath VOCs were assayed by GC/MS in 42 patients hospitalized for suspicion of pulmonary tuberculosis and in 59 healthy controls. Sputum cultures were positive for Mycobacteria in 23/42 and negative in19/42 patients. Breath markers of oxidative stress were increased in all hospitalized patients (p<0.04). Pattern recognition analysis and fuzzy logic analysis of breath VOCs independently distinguished healthy controls from hospitalized patients with 100% sensitivity and 100% specificity. Fuzzy logic analysis identified patients with positive sputum cultures with 100% sensitivity and 100% specificity (95.7% sensitivity and 78.9% specificity on leave-one-out cross-validation); breath VOC markers were similar to those observed in vitro, including naphthalene, 1-methyl- and cyclohexane, 1,4-dimethyl-. Pattern recognition analysis identified patients with positive sputum cultures with 82.6% sensitivity (19/23) and 100% specificity (18/18), employing 12 principal components from 134 breath VOCs. We conclude that volatile biomarkers in breath were sensitive and specific for pulmonary tuberculosis: the breath test distinguished between "sick versus well" i.e. between normal controls and patients hospitalized for suspicion of pulmonary tuberculosis, and between infected versus non-infected patients i.e. between those whose sputum cultures were positive or negative for Mycobacteria.
authors.affiliation: Menssana Research Inc., Fort Lee, NJ 07024, USA. mphillips@menssanaresearch.com
authors.forename: Michael
authors.initials: M
authors.lastname: Phillips
authors.forename: Renee N
authors.initials: RN
authors.lastname: Cataneo
authors.forename: Rany
authors.initials: R
authors.lastname: Condos
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Ring Erickson
authors.forename: Joel
authors.initials: J
authors.lastname: Greenberg
authors.forename: Vincent
authors.initials: V
authors.lastname: La Bombardi
authors.forename: Muhammad I
authors.initials: MI
authors.lastname: Munawar
authors.forename: Olaf
authors.initials: O
authors.lastname: Tietje
mesh_terms: D000328:Adult; D000473:Alkanes / Q000032:analysis; D001555:Benzene Derivatives / Q000032:analysis; D015415:Biomarkers / Q000032:analysis*; D001944:Breath Tests / Q000379:methods*; D001987:Bronchiectasis / Q000175:diagnosis / Q000382:microbiology; D017143:Fuzzy Logic; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D008875:Middle Aged; D015999:Multivariate Analysis; D009169:Mycobacterium tuberculosis / Q000378:metabolism; D009281:Naphthalenes / Q000032:analysis; D018384:Oxidative Stress / Q000502:physiology; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis*
chemical_list: D000473:Alkanes; D001555:Benzene Derivatives; D015415:Biomarkers; D009281:Naphthalenes