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Applied and Environmental Microbiology, October 2005, p. 6086-6095, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6086-6095.2005

Bioaerosol Mass Spectrometry for Rapid Detection of Individual Airborne Mycobacterium tuberculosis H37Ra Particles

Herbert J. Tobias,¹ Millie P. Schafer,² Maurice Pitesky,¹ David P. Fergenson,¹ Joanne Horn,¹ Matthias Frank,¹ and Eric E. Gard^1*

Lawrence Livermore National Laboratory, Livermore, California,¹ National Institute for Occupational Safety and Health, Cincinnati, Ohio²

Received 2 March 2005/ Accepted 22 April 2005

Single-particle laser desorption/ionization time-of-flight mass spectrometry, in the form of bioaerosol mass spectrometry (BAMS), was evaluated as a rapid detector for individual airborne, micron-sized, Mycobacterium tuberculosis H37Ra particles, comprised of a single cell or a small number of clumped cells. The BAMS mass spectral signatures for aerosolized M. tuberculosis H37Ra particles were found to be distinct from M. smegmatis, Bacillus atrophaeus, and B. cereus particles, using a distinct biomarker. This is the first time a potentially unique biomarker was measured in M. tuberculosis H37Ra on a single-cell level. In addition, M. tuberculosis H37Ra and M. smegmatis were aerosolized into a bioaerosol chamber and were sampled and analyzed using BAMS, an aerodynamic particle sizer, a viable Anderson six-stage sampler, and filter cassette samplers that permitted direct counts of cells. In a background-free environment, BAMS was able to sample and detect M. tuberculosis H37Ra at airborne concentrations of >1 M. tuberculosis H37Ra-containing particles/liter of air in 20 min as determined by direct counts of filter cassette-sampled particles, and concentrations of >40 M. tuberculosis H37Ra CFU/liter of air in 1 min as determined by using viable Andersen six-stage samplers. This is a first step toward the development of a rapid, stand-alone airborne M. tuberculosis particle detector for the direct detection of M. tuberculosis bioaerosols generated by an infectious patient. Additional instrumental development is currently under way to make BAMS useful in realistic environmental and respiratory particle backgrounds expected in tuberculosis diagnostic scenarios.

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* Corresponding author. Mailing address: L-452 Lawrence Livermore National Laboratory, 7000 East Ave, Livermore, CA 94550. Phone: (925) 422-0038. Fax: (925) 422-3570. E-mail: gard2{at}llnl.gov.

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Applied and Environmental Microbiology, October 2005, p. 6086-6095, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6086-6095.2005

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