21 December 1999 Detection of biological aerosols by luminescence techniques
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Luciferin-Luciferase (L-L) luminescence techniques were used to successfully measure adenosine triphosphate (ATP) (pg/ml) in concentrated aerosol samples containing either vegetative bacterial cells or spores. Aerosols were collected with wet and dry sampling devices. Evaluation for the presence of total bio-mass from bacterial and non-bacterial sources of ATP was achieved by suspending the collected aerosol samples in phosphate buffered saline (PBS), pipeting a 50-(mu) l aliquot of the PBS suspension into a FiltravetteTM, and then adding bacterial releasing agent (BRA). The sample was then reacted with L-L, and the resulting Relative Luminescence Units (RLU's), indicative of ATP from all sources, were measured. Bacterial cells were enumerated with the additional application of a wash with somatic cell releasing agent (SRA) to remove any interferences and non-bacterial sources of ATP prior to BRA application. This step removes interfering substances and non-bacterial sources of ATP. For spore analysis, an equi-volume sample of the PBS suspension was added to an equi-volume of trypticase soy broth (TSB), incubated at 37 C for 15 minutes, and processed using methods identical to bacterial cell analysis. Using these technique we were able to detect Bacillus subtilin variation niger, formerly known as Bacillus globigii (BG), in aerosol samples at concentrations greater than or equal to 105 colony forming units (CFU) per ml. Results of field and chamber trials show that one can detect the presence of bacterial and non-bacterial sources of ATP. One can also differentiate spore and vegetative bacterial cells. These techniques may be appropriate to situations where the measurement of bacterial aerosols is needed.
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Peter J. Stopa, Darlene Tieman, Phillip A. Coon, Dorothea A. Paterno, Maurice M. Milton, "Detection of biological aerosols by luminescence techniques", Proc. SPIE 3853, Environmental Monitoring and Remediation Technologies II, (21 December 1999); doi: 10.1117/12.372848; https://doi.org/10.1117/12.372848

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