Molecular screening of Clostridium difficile toxins A and B genetic determinants and identification of mutant strains.
Three separate sets of polymerase chain reaction primers were designed to specifically detect the presence of a toxin A gene fragment, a toxin B gene fragment, and the entire toxin B gene. In addition toxin gene fragments that were amplified from well characterized toxic strains were tagged fluorescently and used as hybridization probes to screen C. difficile strains. A survey of 37 toxic strains and 10 non-toxic strains demonstrated that toxic strains normally contain the genetic composition for toxin A and toxin B simultaneously; whereas, non-toxic strains typically did not contain detectable toxin determinants. The only exception found was strain 39, which had the genetic composition for toxins A and B, but was not cytotoxic under the conditions tested.
McMillin
D E
DE
D.E.M. Science Research Institute, Atlanta, GA 30310.
Muldrow
L L
LL
Leggette
S J
SJ
Abdulahi
Y
Y
Ekanemesang
U M
UM
D000818:Animals; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D016360:Clostridioides difficile / Q000032:analysis* / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004768:Enterotoxins; D004910:Erythrocyte Membrane; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D011817:Rabbits
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile