Novel evidence suggesting Clostridium difficile is present in human gut microbiota more frequently than previously suspected.
Prevalence rate of Clostridium difficile in healthy human adults is believed to be very low. Our RT-PCR system using glass powder, which can eliminate PCR inhibitors, detected C. difficile toxin B mRNA in 16 of 30 fecal samples (53.3%) from healthy human adults. In contrast, we failed to detect toxin B in the same fecal samples by PCR using DNA templates extracted with phenol-chloroform. Our results suggest that PCR inhibitors in feces carried through phenol-chloroform extraction procedure might suppress the sensitivity of PCR and that C. difficile is actually present in human gut microbiota more frequently than previously suspected.
Iizuka
Masahiro
M
Department of Internal Medicine, Akita University School of Medicine, Japan.
Konno
Shiho
S
Itou
Hiroaki
H
Chihara
Junichi
J
Toyoshima
Itaru
I
Horie
Yasuo
Y
Sasaki
Kenji
K
Sato
Akiko
A
Shindo
Kenichi
K
Watanabe
Sumio
S
D000328:Adult; D001427:Bacterial Toxins / Q000235:genetics*; D002725:Chloroform; D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology; D005260:Female; D005898:Glass; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D019800:Phenol; D011208:Powders; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013698:Templates, Genetic
D001427:Bacterial Toxins; C488583:Clostridium difficile lethal toxin B; D004269:DNA, Bacterial; D011208:Powders; D019800:Phenol; D002725:Chloroform