A rapid PCR-based DNA test for enterotoxic Bacillus cereus.
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.
Mäntynen
V
V
Department of Applied Chemistry and Microbiology, University of Helsinki, Finland. Vesa.Mantynen@Helsinki.Fi
Lindström
K
K
D001409:Bacillus cereus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis*; D004768:Enterotoxins / Q000235:genetics*; D006460:Hemolysin Proteins; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004768:Enterotoxins; D006460:Hemolysin Proteins; D012336:RNA, Ribosomal, 16S; C083961:hemolysin BL protein, Bacillus