Selection of primers for specific detection of Clostridium botulinum types B and E neurotoxin genes using PCR method.
Improved oligonucleotide primers were designed to flank 370- and 307-bp fragments of the bont genes encoding botulinum neurotoxins types B and E, respectively. Primer specificity was confirmed for reference strains of Clostridium botulinum types B and E for strains representing bacterial species common in food, and for the DNA mixtures of C. botulinum types B and E in the presence of background DNA isolated from cold smoked salmon and ham. The detection limit of template DNAs of C. botulinum types B and E from the DNA mixtures increased from 1 to 0.1 ng by raising annealing temperature from 50 degrees C to 62 degrees C.
Alsallami
A A
AA
Department of Food Chemistry and Technology, Chemical Faculty, Technical University of Gdańsk, Poland.
Kotłowski
R
R
D000818:Animals; D001483:Base Sequence; D003014:Clostridium botulinum / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers; D005516:Food Microbiology; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012474:Salmon / Q000382:microbiology; D012680:Sensitivity and Specificity; D013552:Swine / Q000382:microbiology; D013696:Temperature
D017931:DNA Primers; D009498:Neurotoxins