Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples

Irenge, L M; Durant, J F; Tomaso, H; Pilo, Paola; Olsen, J S; Ramisse, V; Mahillon, J; Gala, J L (2010). Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples. Applied microbiology and biotechnology, 88(5), pp. 1179-92. Heidelberg: Springer 10.1007/s00253-010-2848-0

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A real-time polymerase chain reaction (PCR) assay was developed for rapid identification of Bacillus anthracis in environmental samples. These samples often harbor Bacillus cereus bacteria closely related to B. anthracis, which may hinder its specific identification by resulting in false positive signals. The assay consists of two duplex real-time PCR: the first PCR allows amplification of a sequence specific of the B. cereus group (B. anthracis, B. cereus, Bacillus thuringiensis, Bacillus weihenstephanensis, Bacillus pseudomycoides, and Bacillus mycoides) within the phosphoenolpyruvate/sugar phosphotransferase system I gene and a B. anthracis specific single nucleotide polymorphism within the adenylosuccinate synthetase gene. The second real-time PCR assay targets the lethal factor gene from virulence plasmid pXO1 and the capsule synthesis gene from virulence plasmid pXO2. Specificity of the assay is enhanced by the use of minor groove binding probes and/or locked nucleic acids probes. The assay was validated on 304 bacterial strains including 37 B. anthracis, 67 B. cereus group, 54 strains of non-cereus group Bacillus, and 146 Gram-positive and Gram-negative bacteria strains. The assay was performed on various environmental samples spiked with B. anthracis or B. cereus spores. The assay allowed an accurate identification of B. anthracis in environmental samples. This study provides a rapid and reliable method for improving rapid identification of B. anthracis in field operational conditions.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Pilo, Paola

Subjects:

600 Technology > 630 Agriculture

ISSN:

0175-7598

Publisher:

Springer

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:32

Last Modified:

13 Apr 2017 16:04

Publisher DOI:

10.1007/s00253-010-2848-0

Web of Science ID:

000283095500015

URI:

https://boris.unibe.ch/id/eprint/12353 (FactScience: 218680)

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