Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides differentiation using a PCR-RE technique.
A method was developed to differentiate between Bacillus cereus, Bacillus mycoides and Bacillus thuringiensis using the polymerase chain reaction combined with a restriction endonuclease (PCR-RE) technique. This fast and simple protocol, applied to pure culture strains, was developed using the gyrB DNA sequence, as previously proposed by other authors. Strains from international collections were used to optimize the method which was then applied to the identification of strains isolated from food samples. Amplifications were specific for the B. cereus group. Only Staphylococcus aureus gave the same size PCR product, but it was easily differentiated from strains in the B. cereus group by using restriction analysis, based on digestion with the RsaI, Sau3AI and EcoRI endonucleases. Specific amplifications and good differentiations were obtained using pure strains, suggesting the possibility of using the method described to identify the B. cereus group directly in food samples.
Manzano
Marisa
M
Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università degli Studi di Udine, Italy. marisa.manzano@DSA.uniud.it
Cocolin
Luca
L
Cantoni
Carlo
C
Comi
Giuseppe
G
D001407:Bacillus / Q000145:classification* / Q000302:isolation & purification; D001409:Bacillus cereus / Q000145:classification / Q000302:isolation & purification; D001413:Bacillus thuringiensis / Q000145:classification / Q000302:isolation & purification; D001483:Base Sequence; D027081:DNA Gyrase / Q000235:genetics; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D005504:Food Analysis; D005516:Food Microbiology; D005819:Genetic Markers; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers; D004262:DNA Restriction Enzymes; D027081:DNA Gyrase