A multiplex loop-mediated isothermal amplification assay for rapid detection of Bacillus cereus and Staphylococcus aureus.

Bacillus cereus (B. cereus) and Staphylococcus aureus (S. aureus) are major human food-borne pathogens that may produce a variety of toxins and cause diarrhea, food poisoning, and even death. In order to monitor and prevent the spread of these pathogens, a multiplex loop-mediated isothermal amplification (multi-LAMP) assay was developed to simultaneously and rapidly detect B. cereus and S. aureus. The sensitivity and specificity of the loop-mediated isothermal amplification (LAMP) reactions were determined via electrophoresis. The multi-LAMP showed 100% inclusivity and exclusivity, the sensitivity was 10 fg/μL and was 10 times more sensitive than that of polymerase chain reaction (PCR), the results were consistent with those of conventional PCR assay, and the entire assay should be finished within 40 min. This multi-LAMP assay was confirmed as a rapid and reliable diagnostic technique upon application for clinical samples and food samples. To our knowledge, this is the first study to report the application of multi-LAMP to detect B. cereus and S. aureus.