title: Algorithm combining toxin immunoassay and stool culture for diagnosis of Clostridium difficile infection.
abstract: Enzyme immunoassays (EIA) to detect glutamate dehydrogenase or toxins A (TcdA) and B (TcdB), a cytotoxicity assay, and bacteriologic culture have disadvantages when applied individually to diagnosis of Clostridium difficile infections. Stool specimens (n = 1,596) were subjected to toxin detection via an enzyme-linked fluorescent immunoassay (ELFA; Vidas CDAB assay) and bacteriologic culture for toxigenic C. difficile in a three-step algorithm with additional toxigenic culture. Isolates (n = 163) from ELFA-negative stool specimens were examined via ELFA for toxin production. We amplified tcdA and tcdB from C. difficile isolates and tcdB from stool specimens that were ELFA positive or equivocal and culture negative, and we compared the results to those obtained with the three-step algorithm. More than 26% of stool specimens (419/1,596) were culture positive, yielding 248 isolates (59.2%) with both toxin genes (tcdA- and tcdB-positive isolates), 88 isolates (21.0%) with either tcdA or tcdB, and 83 (19.8%) that had no toxin genes (tcdA- and tcdB-negative isolates). Among 49 (culture-negative/ELFA-positive or -equivocal) stool specimens, 53.1% (26/49) represented tcdB-positive isolates. Therefore, the total number of PCR-positive cases was 362, and 27.1% (98/362) of these were detected through toxigenic culture. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 63.3%, 96.7%, 90.5%, and 92.4% (ELFA alone); 92.8%, 93.3%, 80.2%, and 97.8% (culture); and 70.7%, 91.4%, 95.5%, and 100% (three-step algorithm ELFA and bacterial culture with toxigenic culture), respectively, with culture and PCR for tcdA and tcdB as the standards. Thus, sensitivity and specificity were highest using culture and ELFA, respectively, but we recommend the three-step algorithm comprising EIA to detect both toxins and toxigenic culture for C. difficile as a practical method for achieving better PPV and NPV.
authors.affiliation: Department of Laboratory Medicine, Sanggye Paik Hospital, Inje University, Seoul, South Korea.
authors.forename: Bo-Moon
authors.initials: BM
authors.lastname: Shin
authors.forename: Eun Young
authors.initials: EY
authors.lastname: Kuak
authors.forename: Eun Joo
authors.initials: EJ
authors.lastname: Lee
authors.forename: J Glenn
authors.initials: JG
authors.lastname: Songer
mesh_terms: D000465:Algorithms; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D016360:Clostridioides difficile / Q000254:growth & development / Q000302:isolation & purification*; D004761:Enterocolitis, Pseudomembranous / Q000175:diagnosis*; D004768:Enterotoxins / Q000032:analysis* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005243:Feces / Q000737:chemistry* / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile