title: Novel evidence suggesting Clostridium difficile is present in human gut microbiota more frequently than previously suspected.
abstract: 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.
authors.affiliation: Department of Internal Medicine, Akita University School of Medicine, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Iizuka
authors.forename: Shiho
authors.initials: S
authors.lastname: Konno
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Itou
authors.forename: Junichi
authors.initials: J
authors.lastname: Chihara
authors.forename: Itaru
authors.initials: I
authors.lastname: Toyoshima
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Horie
authors.forename: Kenji
authors.initials: K
authors.lastname: Sasaki
authors.forename: Akiko
authors.initials: A
authors.lastname: Sato
authors.forename: Kenichi
authors.initials: K
authors.lastname: Shindo
authors.forename: Sumio
authors.initials: S
authors.lastname: Watanabe
mesh_terms: 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
chemical_list: D001427:Bacterial Toxins; C488583:Clostridium difficile lethal toxin B; D004269:DNA, Bacterial; D011208:Powders; D019800:Phenol; D002725:Chloroform