Differentiation of Bacillus anthracis from Other Bacillus cereus Group Bacteria with the PCR

Henderson, Ian; Duggleby, Clive J.; Turnbull, P. C. B.

doi:10.1099/00207713-44-1-99

Cited by 89 publications

(72 citation statements)

References 22 publications

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“…M13-PCR typing is based on random PCR amplification of DNA fragments using an Escherichia coli phage M13-based primer. It was adapted from Henderson et al (1994) using the sequence-specific primer PM13 (for primer sequences see Table 2). DNA was prepared as described previously (Guinebretiere et al, 2002).…”

Section: Methodsmentioning

confidence: 99%

Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains

Svensson²,

et al. 2005

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An in-depth polyphasic approach was applied to study the population structure of the human pathogen Bacillus cereus. To assess the intraspecific biodiversity of this species, which is the causative agent of gastrointestinal diseases, a total of 90 isolates from diverse geographical origin were studied by genetic [M13-PCR, random amplification of polymorphic DNA (RAPD), multilocus sequence typing (MLST)] and phenetic [Fourier transform Infrared (FTIR), protein profiling, biochemical assays] methods. The strain set included clinical strains, isolates from food remnants connected to outbreaks, as well as isolates from diverse food environments with a well documented strain history. The phenotypic and genotypic analysis of the compiled panel of strains illustrated a considerable diversity among B. cereus connected to diarrhoeal syndrome and other non-emetic food strains, but a very low diversity among emetic isolates. Using all typing methods, cluster analysis revealed a single, distinct cluster of emetic B. cereus strains. The isolates belonging to this cluster were neither able to degrade starch nor could they ferment salicin; they did not possess the genes encoding haemolysin BL (Hbl) and showed only weak or no haemolysis. In contrast, haemolytic-enterotoxin-producing B. cereus strains showed a high degree of heterogeneity and were scattered over different clusters when different typing methods were applied. These data provide evidence for a clonal population structure of cereulide-producing emetic B. cereus and indicate that emetic strains represent a highly clonal complex within a potentially panmictic or weakly clonal background population structure of the species. It may have originated only recently through acquisition of specific virulence factors such as the cereulide synthetase gene.Abbreviations: FTIR, Fourier transform Infrared; Hbl, haemolysin BL; MLST, multilocus sequence typing; Nhe, non-haemolytic enterotoxin; RAPD, random amplification of polymorphic DNA.The GenBank/EMBL/DDBJ accession numbers for the sequences of the internal gene fragments used for MLST and for the sporulation stage III AB genes reported in this paper are AY762151-AY762213 and AY578317-AY578349, respectively.

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Section: Methodsmentioning

confidence: 99%

Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains

Svensson²,

et al. 2005

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“…It has been applied frequently to distinguish subgroups of closely related organisms including those showing a high degree of sequence conservation among isolates, as in the case of Bacillus anthracis (Henderson et al, 1994). In this study, RAPD-PCR was used to fingerprint the whole genome of B. cereus strains NCIB 8122 and MP01.…”

Section: Strain Mp01 Is a Fliy Null Mutantmentioning

confidence: 99%

Swarming motility in Bacillus cereus and characterization of a fliY mutant impaired in swarm cell differentiation The EMBL accession number for the sequence reported in this paper is Y08031.

et al. 2002

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This report describes a new behavioural response of Bacillus cereus that consists of a surface-induced differentiation of elongated and hyperflagellated swarm cells exhibiting the ability to move collectively across the surface of the medium. The discovery of swarming motility in B. cereus paralleled the isolation of a spontaneous non-swarming mutant that was found to carry a deletion of fliY, the homologue of which, in Bacillus subtilis, encodes an essential component of the flagellar motor-switch complex. However, in contrast to B. subtilis, the fliY mutant of B. cereus was flagellated and motile, thus suggesting a different role for FliY in this organism. The B. cereus mutant was completely deficient in chemotaxis and in the secretion of the L 2 component of the tripartite pore-forming necrotizing toxin, haemolysin BL, which was produced exclusively by the wild-type strain during swarm-cell differentiation. All the defects in the fliY mutant of B. cereus could be complemented by a plasmid harbouring the B. cereus fliY gene. These results demonstrate that the activity of fliY is required for swarming and chemotaxis in B. cereus, and suggest that swarm-cell differentiation is coupled with virulence in this organism.

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“…It is a member of the B. cereus group including B. cereus, B. mycoides, and B. thuringiensis. This is one of the most taxonomically ambiguous groups of bacilli (17,28). These species have been shown indistinguishable in the comparison of DNA-DNA hybridization, 16S-23S rDNA sequences, and gyrB-gyrA intergenic spacer regions (1,7,18,22,34).…”

mentioning

confidence: 99%

Sensitive and Rapid Quantitative Detection of Anthrax Spores Isolated from Soil Samples by Real‐Time PCR

Ryu

Lee

Yoo

et al. 2003

Microbiology and Immunology

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Quantitative analysis of anthrax spores from environmental samples is essential for accurate detection and risk assessment since Bacillus anthracis spores have been shown to be one of the most effective biological weapons. Using TaqMan real-time PCR, specific primers and probes were designed for the identification of pathogenic B. anthracis strains from pag gene and cap gene on two plasmids, pXO1 and pXO2, as well as a sap gene encoded on the S-layer. To select the appropriate lysis method of anthrax spore from environmental samples, several heat treatments and germination methods were evaluated with multiplex-PCR. Among them, heat treatment of samples suspended with sucrose plus non-ionic detergent was considered an effective spore disruption method because it detected up to 10 5 spores/g soil by multiplex-PCR. Serial dilutions of B. anthracis DNA and spore were detected up to a level of 0.1 ng/l and 10 spores/ml, respectively, at the correlation coefficient of 0.99 by real-time PCR. Quantitative analysis of anthrax spore could be obtained from the comparison between CT value and serial dilutions of soil sample at the correlation coefficient of 0.99. Additionally, spores added to soil samples were detected up to 10 4 spores/g soil within 3 hr by real-time PCR. As a consequence, we established a rapid and accurate detection system for environmental anthrax spores using real-time PCR, avoiding time and labor-consuming preparation steps such as enrichment culturing and DNA preparation.

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Differentiation of Bacillus anthracis from Other Bacillus cereus Group Bacteria with the PCR

Cited by 89 publications

References 22 publications

Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains

Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains

Swarming motility in Bacillus cereus and characterization of a fliY mutant impaired in swarm cell differentiation The EMBL accession number for the sequence reported in this paper is Y08031.

Sensitive and Rapid Quantitative Detection of Anthrax Spores Isolated from Soil Samples by Real‐Time PCR

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