Detection of bioterror agents in air samples using real-time PCR

• Published in: Journal of applied microbiology Vol. 105; no. 2; pp. 351 - 358
• Main Authors: Fykse, E.M, Langseth, B, Olsen, J.S, Skogan, G, Blatny, J.M
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.08.2008; Blackwell Publishing Ltd; Blackwell Science
• Subjects: Air Microbiology; airborne bacteria; airborne microorganisms; Bacteria - genetics; Bacteria - isolation & purification; bioaerosol; Biological and medical sciences; bioterror agents; Bioterrorism; Environmental Monitoring - methods; Fundamental and applied biological sciences. Psychology; Microbiology; real-time PCR; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; Norway; Applied microbiology; Sample; bioaerosol; bioterror agents; Air; real-time PCR; Bioterrorism; Real time; Polymerase chain reaction; Analysis method; airborne bacteria; Bacteria; Detection
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2008.03750.x

To use real-time PCR for the detection of bacterial bioterror agents in a liquid air sample containing potential airborne interferences, including bacteria, without the need for DNA extraction. Bacteria in air were isolated after passive sedimentation onto R2A agar plates and characterized by 16S rRNA sequencing. Real-time PCR was used to identify different bacterial bioterror agents in an artificial air sample consisting of a liquid air sample and a mixture of miscellaneous airborne bacteria showing different colony morphology on R2A agar plates. No time-consuming DNA extraction was performed. Specifically designed fluorescent hybridization probes were used for identification. Fourteen different bacterial genera were classified by 16S rRNA gene sequencing of selected bacterial colonies showing growth on R2A agar plates. Real-time PCR amplification of all the bacterial bioterror agents was successfully obtained in the artificial air sample containing commonly found airborne bacteria and other potential airborne PCR interferences. Bacterial bioterror agents can be detected within 1 h in a liquid air sample containing a variety of commonly found airborne bacteria using real-time PCR. Airborne viable bacteria at Kjeller (Norway) were classified to the genera level using 16S rRNA gene sequencing.