Effects of inactivation methods on the analysis of Bacillus atrophaeus endospores using real-time PCR and MALDI-TOF-MS

verfasst von: Steven R. Talbot, Heiko Russmann, Stefan Köhne, Bärbel Niederwöhrmeier, Gudrun Grote, Thomas Scheper
Abstract: A wide range of analytical methods are available for the detection and identification of biological warfare agents. These technologies are often hampered in their performance when the inactivated samples are analyzed. To work with pathogens outside of biosafety level 3 laboratories, a complete inactivation is mandatory when appropriate protection equipment is unavailable. When methods of inactivation are used, the detection of bacteria becomes more difficult. In contrast to measuring viable organisms, inactivation steps can have a massive impact on the intrinsic cellular information. This study examined the effects of autoclaving and chemical inactivation methods on Bacillus spores using biological warfare detection setups like real-time PCR and MALDI-TOF-MS. Here, the inactivation of Bacillus atrophaeus spores with formaldehyde, which is a suggested model for biological warfare spore agents, was compared with other inactivation reagents like WofasterilsE400, a commercially available decontaminant based on peroxyacetic acid. With WofasterilsE400 the critical factor of inactivation time was reduced to about 15 min and a limit of detection of 8500 spores by PCR was still measurable using five-times-washed spores. It has also been shown that MALDITOF-MS peak information can be hampered by inactivation methods.
Organisationseinheit(en): Institut für Technische Chemie
Externe Organisation(en): Wehrwissenschaftliches Institut Für Schutztechnologien - ABC-Schutz (WIS)
Typ: Artikel
Journal: Engineering in life sciences
Band: 10
Seiten: 109-120
Anzahl der Seiten: 12
ISSN: 1618-0240
Publikationsdatum: 12.04.2010
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Biotechnologie, Environmental engineering, Bioengineering
Elektronische Version(en): https://doi.org/10.1002/elsc.200800078 (Zugang: Unbekannt)

BibTeX

@article{a228853603dd4a4986e9396587658baa,
title = "Effects of inactivation methods on the analysis of Bacillus atrophaeus endospores using real-time PCR and MALDI-TOF-MS",
abstract = "A wide range of analytical methods are available for the detection and identification of biological warfare agents. These technologies are often hampered in their performance when the inactivated samples are analyzed. To work with pathogens outside of biosafety level 3 laboratories, a complete inactivation is mandatory when appropriate protection equipment is unavailable. When methods of inactivation are used, the detection of bacteria becomes more difficult. In contrast to measuring viable organisms, inactivation steps can have a massive impact on the intrinsic cellular information. This study examined the effects of autoclaving and chemical inactivation methods on Bacillus spores using biological warfare detection setups like real-time PCR and MALDI-TOF-MS. Here, the inactivation of Bacillus atrophaeus spores with formaldehyde, which is a suggested model for biological warfare spore agents, was compared with other inactivation reagents like WofasterilsE400, a commercially available decontaminant based on peroxyacetic acid. With WofasterilsE400 the critical factor of inactivation time was reduced to about 15 min and a limit of detection of 8500 spores by PCR was still measurable using five-times-washed spores. It has also been shown that MALDITOF-MS peak information can be hampered by inactivation methods.",
keywords = "Biological warfare agents, BSL3 bacteria, Endospores, Fast detection systems, Inactivation",
author = "Talbot, {Steven R.} and Heiko Russmann and Stefan K{\"o}hne and B{\"a}rbel Niederw{\"o}hrmeier and Gudrun Grote and Thomas Scheper",
year = "2010",
month = apr,
day = "12",
doi = "10.1002/elsc.200800078",
language = "English",
volume = "10",
pages = "109--120",
journal = "Engineering in life sciences",
issn = "1618-0240",
publisher = "Wiley-VCH Verlag",
number = "2",
}