Rapid Detection of Bacillus cereus Using Cross-Priming Amplification

Abstract

ABSTRACT Bacillus cereus is a spore-forming gastrointestinal pathogen that can cause life-threatening diseases. Here, a simple and effective assay to detect B. cereus was developed, using cross-priming amplification (CPA). Amplicons were detected using disposable cartridges that contained nucleic acid detection strips. The sensitivity of CPA assay for B. cereus was assessed using serial dilutions of genomic DNA, which indicated a detection limit of 3.6 × 101 CFU/mL. No cross-reactions were detected when genomic DNA extracted from 12 different B. cereus strains and 20 other bacterial foodborne strains were tested, suggesting that the assay is highly specific. Finally, we evaluated the practical applications of the CPA assay for the detection of B. cereus in 150 food samples and found that its sensitivity and specificity, compared with real-time PCR, were approximately 98.18 and 100%, respectively. In conclusion, CPA combined with nucleic acid detection strips is easy to perform, requires simple equipment, and offers highly specific and sensitive B. cereus detection.