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Appl Environ Microbiol, May 1998, p. 1634-1639, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Rapid PCR-Based DNA Test for Enterotoxic Bacillus cereus

Vesa Mäntynen* and Kristina Lindström

Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Biocenter, FIN-00014 University of Helsinki, Finland

Received 19 November 1997/Accepted 23 February 1998

The occurrence of DNA sequences encoding the hemolysin HblA complex and Bacillus cereus enterotoxin BceT, which have recently been confirmed as enterotoxins, was studied in Bacillus spp. To amplify these DNA sequences, PCR primer systems for the B component of hblA and for bceT DNA sequences were developed. The results from the amplification of hblA sequences correlated well with results obtained with the B. cereus enterotoxin (diarrheal type) test kit (RPLA kit), but not with the results of the Bacillus diarrheal enterotoxin visual immunoassay (BDE kit). Except for two thermophilic strains, all strains that were positive in PCR amplification assays with the hblA primers were also positive when tested with the RPLA kit. The hblA DNA sequence was found in 33 strains, and these strains were closely related according to 16S rDNA-RFLP analysis, except B. pasteurii. In PCR amplifications with the bceT primers only the model strain gave a positive signal. It is concluded that screening of the hemolysin HblA complex by the PCR method allows faster detection of enterotoxin production than does testing with the RPLA enterotoxin kit.

* Corresponding author. Mailing address: Department of Applied Chemistry and Microbiology, Division of Microbiology, University of Helsinki, Biocenter, P.O. Box 56, FIN-00014 University of Helsinki, Finland. Phone: 358-9-70859277. Fax: 358-9-70859322. E-mail: Vesa.Mantynen{at}Helsinki.Fi.

Appl Environ Microbiol, May 1998, p. 1634-1639, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.