Establishment of ERIC-PCR Molecular Typing Method for Bacillus cereus
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Abstract:
An efficient enterobacterial repetitive intergenic consensus sequence (ERIC-PCR) molecular typing method was established for Bacillus cereus. Based on the template of B. cereus CMCC 63303 genomic DNA, the ERIC-PCR amplification system in four levels of five factors (DNA template, Mg2+, dNTP, Taq DNA polymerase and primer) was optimized by orthogonal design L16(45) .Then the effects of annealing temperature on the amplification system was discussed through single factor experiment and the optimal reaction condition were determined. Fifty B. cereus strains were typed using the optimal reaction system and cluster analysis. The ERIC-PCR results exhibited better discriminative results in molecular typing with discrimination index of 0.996. B. cereus strains were grouped into 47 types. The ERIC-PCR system was an efficient method for typing and tracking analyses. It was suitable for genetic diversity analysis of foodborne B. cereus strains.
