Q-178. Rapid Detection for Bioaerosol Monitoring Using ATP Bioluminescence, qRT-PCR or Real Time Mediated Amplification

K. S. Souza, M. Aysola, S. Rigby, J. Broe;
Millipore Corp., Bedford, MA.

Background: Environmental monitoring for viable airborne microorganisms is often performed by collection and enumeration of microorganisms with air impaction on agar media plates requiring up to seven days for results. Rapid feedback on the status of environmental contamination would enable a prompt response from pharmaceutical or hospital personnel. Methods: Air samples were collected into a cyclone liquid collection device followed by filtration on a membrane filter and rapid enumeration by ATP bioluminescence. Multiple experiments were designed with either air samples or spiked representative microorganisms including Pseudomonas aeruginosa to examine a variety of factors including the effect of six different buffers on microorganism recovery and the effect of vortex on microorganism recovery over time. Additionally, specific nucleic acid detection was also demonstrated with the same spiked sample using Real Time Transcription Mediated Amplification (TMA) and quantitative Reverse Transcriptase Polymerase chain reaction (qRT-PCR). Results: A comparison of recovery of microorganisms from air demonstrated that enumeration by either agar culture media or ATP bioluminescence were equivalent in sensitivity however the ATP bioluminescence method provided results in 24 hours as compared to five to seven days for the agar media plates. The impact of collection liquid type on recovery of microorganisms ranged from 60 to 100 percent. Buffered or physiological salt solutions provided the highest recoveries. The results with P. aeruginosa spiked into collection liquids and subsequently detected by Real Time TMA and PCR showed detection sensitivity from 10 to 1000 cfu per collection volume in four hours without inhibition by the collection method. Conclusion: ATP bioluminescence on membrane filters can be used as a rapid, sensitive, quantitative alternative method for monitoring air quality and can be coupled with either PCR or Real Time TMA for specific microorganism detection.

146/Q. Biodegradation of Lignin and Hydrocarbons

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