Detection of non-emetic and emetic Bacillus cereus by propidium monoazide multiplex PCR (PMA-mPCR) with internal amplification control

摘要: Abstract Bacillus cereus is an etiological agent of food-borne disease that can cause a type emesis. To develop sensitive and reliable diagnosis technique for detecting all the species B. cereus group, specific primers were designed to target recently discovered part cereulide synthetase gene ( cesB ) emetic 16S rRNA non-emetic . detect PCR signals only from viable cells, propidium monoazide (PMA) was selected eliminate false-positive results. In addition, internal amplification control (IAC) applied meet diagnostic multiplex requirements will prevent occurrence false-negative The inclusivity exclusivity mPCR assay estimated using panel 100 strains, including 2 , 77 21 non- strains. limit detection (LOD) dead without PMA treatment in pure bacteria culture 4.0 × 10  CFU/mL, as low 7.5 × 10 0  CFU/mL treatment, 1 with treatment. spiked food produce detected specifically sensitively at 1.0 × 10 3  CFU/g which lowest concentration detected. This novel PMA-mPCR-IAC rapid reliable, providing efficient tool promising application monitoring samples.