An alternative real-time PCR method to detect the Bacillus cereus group in naturally contaminated food gelatine: a comparison study.
OBJECTIVE
Comparison of an internally controlled real-time PCR assay with the standard plate-based assay (ISO 21871) for the detection of Bacillus cereus group cells in gelatine.


RESULTS
A comprehensive TaqMan probe was designed allowing the real-time PCR assay to be fully inclusive and exclusive. An internal amplification control was designed and implemented at 500 copies per reaction without impact on target detection. Specific and selective detection of target cells was achieved with a quick and simple DNA preparation procedure. No significant difference (kappa = 0.99) was observed between the performance of the real-time PCR and the standard plate-based method on naturally contaminated gelatines (n = 197). Relative accuracy, relative sensitivity and relative specificity were > or =99%.


CONCLUSIONS
The real-time PCR assay is a valid alternative of the standard plate-based assay.


CONCLUSIONS
The real-time PCR assay decreased the time between sample collection and result from 2 days to 2 h, while analysis cost did not increase. The gelatine-producing industry can ensure gelatine safety and quality in a much faster way.
Reekmans
R
R
Laboratory of Microbiology, Department of Biochemistry, Physiology and Microbiology, Ghent University, K.L. Ledeganckstraat 35, Ghent, Belgium. Rieka.Reekmans@UGent.be
Stevens
P
P
Vervust
T
T
De Vos
P
P
D001409:Bacillus cereus / Q000302:isolation & purification*; D005516:Food Microbiology; D005780:Gelatin; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity
D015345:Oligonucleotide Probes; D005780:Gelatin