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

Abstract

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.

Microbiology and atmospheric processes: biological, physical and chemical characterization of aerosol particles

“…Nevertheless, by the analysis of fragment ions that are specific to distinct microorganisms, the types of viruses, bacteria, and fungi present in the air can be assessed on an individual-particle basis. The analysis of biomarker ions forms the basis of bioaerosol mass spectrometry (BAMS), a new and highly efficient method for the detection of both spores and vegetative cells of airborne pathogens and bioterrorism agents (Russell et al, 2004Tobias et al, 2005;van Wuijckhuijse et al, 2005;Beddows and Telle, 2005). AMS is now a widely used single-particle method in aerosol science, with unique capabilities for obtaining species-specific data from biological particles.…”

Parameters contributing to efficient ion generation in aerosol MALDI mass spectrometry

“…T he Bioaerosol Mass Spectrometry (BAMS) system was developed for national security and public health applications including the real-time, in situ detection of airborne pathogens [1] and biowarfare agents [2,3]. It has the ability to distinguish Bacillus spores from vegetative cells on a particle by particle basis using ion peaks under m/z 300 [4], and has demonstrated preliminary species level differentiation of individual Bacillus spores using low mass biomarker peaks [5,6].…”

Identification of High Explosives Using Single-Particle Aerosol Mass Spectrometry

“…37 Dale et al demonstrated the detection of 2,4,6-trinitrotoluene (TNT) adsorbed on silicon carbide beads using a single-particle quadrupole ion trap mass spectrometer. 38 Time-of-flight SPAMS at Lawrence Livermore National Laboratory (LLNL) in the form of bioaerosol mass spectrometry (BAMS) has been applied to the detection of various species of Bacillus spores, [39][40][41][42][43][44][45] viruses, 39 toxin simulants, 39 fungi, 41 and Mycobacteria, 45 with and without the use of a chemical matrix during ionization. Matrix-free systems are ideal for real-time and on-line detection systems, as no sample preparation and thus very few reagents are needed for the analysis.…”