Simultaneous detection and identification of Bacillus cereus group bacteria using multiplex PCR.
Bacillus cereus group bacteria share a significant degree of genetic similarity. Thus, to differentiate and identify the Bacillus cereus group efficiently, a multiplex PCR method using the gyrB and groEL genes as diagnostic markers is suggested for simultaneous detection. The assay yielded a 400 bp amplicon for the groEL gene from all the B. cereus group bacteria, and a 253 bp amplicon from B. anthracis, 475 bp amplicon from B. cereus, 299 bp amplicon from B. thuringiensis, and 604 bp amplicon from B. mycoides for the gyrB gene. No nonspecific amplicons were observed with the DNA from 29 other pathogenic bacteria. The specificity and sensitivity of the B. cereus group identification using this multiplex PCR assay were evaluated with different kinds of food samples. In conclusion, the proposed multiplex PCR is a reliable, simple, rapid, and efficient method for the simultaneous identification of B. cereus group bacteria from food samples in a single tube.
Park
Si-Hong
SH
Institute of Life Sciences & Resources, Graduate School of Biotechnology, Kyung Hee University, Suwon 449-701, Korea.
Kim
Hyun-Joong
HJ
Kim
Jae-Hwan
JH
Kim
Tae-Woon
TW
Kim
Hae-Yeong
HY
D001408:Bacillus anthracis / Q000235:genetics*; D001409:Bacillus cereus / Q000235:genetics*; D001413:Bacillus thuringiensis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005516:Food Microbiology; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D012275:Oryza / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014675:Vegetables / Q000382:microbiology
D017931:DNA Primers; D004269:DNA, Bacterial