Detection of Microbial Cells in Aerosols Using Nucleic Acid Probes

Alexander Neef, Rudolf Amann, Karl Heinz Schleifer

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

Non-radioactive hybridization methods were evaluated for the identification of microorganisms in mixed bioaerosols. A cultivation-dependent method, colony hybridization, was compared to a direct, cultivation-independent approach, whole cell hybridization with fluorescently labeled oligonucleotides. After sampling of the aerosols by filtration, special processing of filters (cells) preceded hybridization with fluorescently, digoxigenin- or enzyme-labeled oligonuculeotide probes. Group, genus, or species affiliation of collected cells was analyzed with rRNA-targeted probes. Using nucleic acid probes directed against the multiple cloning site, plasmid bearing Escherichia coli colonies could be differentiated from wild-type colonies. The microbial composition of aerosols ranging from less than one to greater than 109 cells/m3 air could be analyzed with appropriate hybridization formats: whole cell hybridization was only applicable to dense aerosols, colony hybridization yielded best results with lower concentrations. After a short incubation period (several hours), a combination of both formats could be used to rapidly determine the fraction of culturable cells within a bioaerosol. When applying these techniques for the monitoring of aerosols generated by standard microbiological laboratory procedures, low concentrations of airborne Escherichia coli cells (1–450 m−3) could be detected. Compared to conventional air monitoring techniques, hybridization with nucleic acid probes should allow more rapid and reliable detection of airborne microorganisms including genetic engineered microorganisms.

Original languageEnglish
Pages (from-to)113-122
Number of pages10
JournalSystematic and Applied Microbiology
Volume18
Issue number1
DOIs
StatePublished - 1995

Keywords

  • Bioaerosol
  • Colony hybridization
  • Filtration sampling
  • Microorganisms
  • Viable-dead differentiation
  • Whole cell hybridization
  • rRNA-targeted oligonucleotides

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