Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Jan;61(1):98–102. doi: 10.1128/aem.61.1.98-102.1995

Specific oligonucleotide primers for detection of lecithinase-positive Bacillus spp. by PCR.

H Schraft 1, M W Griffiths 1
PMCID: PMC167265  PMID: 7887632

Abstract

An assay based on the PCR has been developed to facilitate detection and identification of Bacillus cereus in foods. Three primers for the PCR have been designed within the sequence for cereolysin AB, a cytolytic determinant that encodes lecithin-hydrolyzing and hemolytic activities of B. cereus. With the PCR and hybridization, the specificity of the primers was tested with 39 isolates of the B. cereus group, with 17 other Bacillus spp., and with 21 non-Bacillus strains. Results demonstrate a high specificity of the three oligonucleotides for isolates of the B. cereus group. With a combined PCR-hybridization assay, the detection limit for B. cereus in artificially contaminated milk was 1 CFU/ml of milk.

Full Text

The Full Text of this article is available as a PDF (277.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ash C., Farrow J. A., Dorsch M., Stackebrandt E., Collins M. D. Comparative analysis of Bacillus anthracis, Bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16S rRNA. Int J Syst Bacteriol. 1991 Jul;41(3):343–346. doi: 10.1099/00207713-41-3-343. [DOI] [PubMed] [Google Scholar]
  2. Benson D., Lipman D. J., Ostell J. GenBank. Nucleic Acids Res. 1993 Jul 1;21(13):2963–2965. doi: 10.1093/nar/21.13.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Darrasse A., Priou S., Kotoujansky A., Bertheau Y. PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases. Appl Environ Microbiol. 1994 May;60(5):1437–1443. doi: 10.1128/aem.60.5.1437-1443.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Flemming C. A., Leung K. T., Lee H., Trevors J. T., Greer C. W. Survival of lux-lac-marked biosurfactant-producing Pseudomonas aeruginosa UG2L in soil monitored by nonselective plating and PCR. Appl Environ Microbiol. 1994 May;60(5):1606–1613. doi: 10.1128/aem.60.5.1606-1613.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Furrer B., Candrian U., Hoefelein C., Luethy J. Detection and identification of Listeria monocytogenes in cooked sausage products and in milk by in vitro amplification of haemolysin gene fragments. J Appl Bacteriol. 1991 May;70(5):372–379. doi: 10.1111/j.1365-2672.1991.tb02951.x. [DOI] [PubMed] [Google Scholar]
  6. Gavrilenko I. V., Baida G. E., Karpov A. V., Kuz'min N. P. Nukleotidnaia posledovatel'nost' genov fosfolipazy C i sfingomielinazy Bacillus cereus BKM-B164. Bioorg Khim. 1993 Jan;19(1):133–138. [PubMed] [Google Scholar]
  7. Gilmore M. S., Cruz-Rodz A. L., Leimeister-Wächter M., Kreft J., Goebel W. A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage. J Bacteriol. 1989 Feb;171(2):744–753. doi: 10.1128/jb.171.2.744-753.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goh S. H., Byrne S. K., Zhang J. L., Chow A. W. Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. J Clin Microbiol. 1992 Jul;30(7):1642–1645. doi: 10.1128/jcm.30.7.1642-1645.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Johansen T., Haugli F. B., Ikezawa H., Little C. Bacillus cereus strain SE-1: nucleotide sequence of the sphingomyelinase C gene. Nucleic Acids Res. 1988 Nov 11;16(21):10370–10370. doi: 10.1093/nar/16.21.10370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johansen T., Holm T., Guddal P. H., Sletten K., Haugli F. B., Little C. Cloning and sequencing of the gene encoding the phosphatidylcholine-preferring phospholipase C of Bacillus cereus. Gene. 1988 May 30;65(2):293–304. doi: 10.1016/0378-1119(88)90466-0. [DOI] [PubMed] [Google Scholar]
  11. Leite R. P., Jr, Egel D. S., Stall R. E. Genetic analysis of hrp-related DNA sequences of Xanthomonas campestris strains causing diseases of citrus. Appl Environ Microbiol. 1994 Apr;60(4):1078–1086. doi: 10.1128/aem.60.4.1078-1086.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Leslie D., Fairweather N., Pickard D., Dougan G., Kehoe M. Phospholipase C and haemolytic activities of Clostridium perfringens alpha-toxin cloned in Escherichia coli: sequence and homology with a Bacillus cereus phospholipase C. Mol Microbiol. 1989 Mar;3(3):383–392. doi: 10.1111/j.1365-2958.1989.tb00183.x. [DOI] [PubMed] [Google Scholar]
  13. Yamada A., Tsukagoshi N., Udaka S., Sasaki T., Makino S., Nakamura S., Little C., Tomita M., Ikezawa H. Nucleotide sequence and expression in Escherichia coli of the gene coding for sphingomyelinase of Bacillus cereus. Eur J Biochem. 1988 Aug 1;175(2):213–220. doi: 10.1111/j.1432-1033.1988.tb14186.x. [DOI] [PubMed] [Google Scholar]
  14. van Netten P., Kramer J. M. Media for the detection and enumeration of Bacillus cereus in foods: a review. Int J Food Microbiol. 1992 Oct;17(2):85–99. doi: 10.1016/0168-1605(92)90108-f. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES