Comparison of multiplex PCR, enzyme immunoassay and cell culture methods for the detection of enterotoxinogenic Bacillus cereus.
Fast and reliable methods are needed for the detection of pathogenic Bacillus cereus which should provide consistent results. Therefore, we tested a panel of 176 strains, including B. cereus strains, B. cereus group strains and other Bacillus spp. with polymerase chain reaction, immunoassays and cytotoxicity tests and assessed the consistency of the results. A screening multiplex PCR for the detection of hbl, nhe, ces and cytK1 as well as two multiplex PCRs for the differentiation of Hbl genes (hblC, hblD, hblA) and Nhe genes (nheA, nheB, nheC) was applied. All PCRs included an internal amplification control. Component specific antibody based immunoassays were used for the detection of the three components of Hbl and Nhe and the overall cytotoxicity to Vero cells and HEp-2 cells was checked. An overall excellent correlation was obtained for the results of the three, methodically independent assays and no false-negative PCR results were seen for any of the strains tested positive in immunoassays and cytotoxicity tests. The three multiplex PCRs proved to be a facile method for the identification of enterotoxinogenic B. cereus isolates.
Wehrle
Esther
E
Department of Veterinary Sciences, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Oberschleissheim, Germany.
Moravek
Maximilian
M
Dietrich
Richard
R
Bürk
Christine
C
Didier
Andrea
A
Märtlbauer
Erwin
E
D001409:Bacillus cereus / Q000235:genetics / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D001427:Bacterial Toxins; D004768:Enterotoxins