title: Evaluation of different methods for detection of Clostridium difficile toxins in Poland.
abstract: The aim of this study was to compare different methods for C. difficile toxins detection. Fifty three stool samples taken from patients with antibiotic-associated diarrhoea were studied. TCD toxin A EIA (Becton Dickinson, USA), Tox A/B ELISA test (TechLab, USA), cytotoxicity and neutralization assay on McCoy cells and PCR for detection of both toxin A and B genes were performed in vivo (in stool samples) and in vitro (in isolated strains). Reference toxigenic and nontoxigenic and two Japanese toxin A-negative and toxin B-positive C. difficile strains were used as a controls. TCD toxin A EIA detected in vivo only 19 positive samples. Tox A/B test detected 52 positive samples out of 53 studied. All 53 stool samples were C. difficile culture positive (53 strains were cultured). Toxin B was detected in 52 strain-supernatants and in all controls (except the nontoxigenic one). Both toxin A and B genes were detected by PCR in all 53 isolated strains, Japanese and reference strain (except the nontoxigenic one). In vitro toxin A was detected by TCD toxin A EIA in 42 strains. These results were compared with those obtained in Tox A/B ELISA test. We observed 52 positive strains. Toxigenic reference strain and two Japanese toxA(-)/toxB(+) strains were also positive. Only 2 negative results were obtained with the nontoxigenic reference strain and unique nontoxigenic isolated strain. Tox A/B ELISA test seems to be the best for detection of C. difficile toxins in vivo and in vitro. Test avoids the false-negative results in the case of presence of toxin A-negative and toxin B-positive strain.
authors.affiliation: Department of Medical Microbiology, Medical University of Warsaw, Poland. gmartir@ib.amwaw.edu.pl
authors.forename: G
authors.initials: G
authors.lastname: Martirosian
authors.forename: H
authors.initials: H
authors.lastname: Pituch
authors.forename: P
authors.initials: P
authors.lastname: Obuch-Woszczatyński
authors.forename: G
authors.initials: G
authors.lastname: Rouyan
authors.forename: F
authors.initials: F
authors.lastname: Meisel-Mikołajczyk
mesh_terms: D000900:Anti-Bacterial Agents / Q000009:adverse effects*; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D001431:Bacteriological Techniques; D016360:Clostridioides difficile / Q000302:isolation & purification* / Q000378:metabolism; D003967:Diarrhea / Q000382:microbiology; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology*; D004768:Enterotoxins / Q000032:analysis* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005243:Feces / Q000382:microbiology; D006801:Humans; D007124:Immunoenzyme Techniques; D011044:Poland; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile