title: A Diagnostic Algorithm for the Detection of Clostridium difficile-Associated Diarrhea.
abstract: BACKGROUND
  Clostridium difficile is a common cause of hospital-acquired diarrhea, which is usually associated with previous antibiotic use. The clinical manifestations of C. difficile infection (CDI) may range from mild diarrhea to fulminant colitis. Clostridium difficile should be considered in diarrhea cases with a history of antibiotic use within the last 8 weeks (community-associated CDI) or with a hospital stay of at least 3 days, regardless of the duration of antibiotic use (hospital-acquired CDI).
  
  
  OBJECTIVE
  This study investigated the frequency of CDI in diarrheic patients and evaluated the efficacy of the triple diagnostic algorithm that is proposed here for C. difficile detection.
  
  
  METHODS
  Cross-sectional study.
  
  
  METHODS
  In this study, we compared three methods currently employed for C. difficile detection using 95 patient stool samples: an enzyme immunoassay (EIA) for toxin A/B (C. diff Toxin A+B; Diagnostic Automation Inc.; Calabasas, CA, USA), an EIA for glutamate dehydrogenase (GDH) (C. DIFF CHEK-60TM, TechLab Inc.; Blacksburg, VA, USA), and a polymerase chain reaction (PCR)-based assay (GeneXpert(®) C. difficile; Cepheid, Sunnyvale, CA, USA) that detects C. difficile toxin genes and conventional methods as well. In this study, 50.5% of the patients were male, 50 patients were outpatients, 32 were from inpatient clinics and 13 patients were from the intensive care unit.
  
  
  RESULTS
  Of the 95 stool samples tested for GDH, 28 were positive. Six samples were positive by PCR, while nine samples were positive for toxin A/B. The hypervirulent strain NAP-1 and binary toxin was not detected. The rate of occurrence of toxigenic C. difficile was 5.1% in the samples. Cefaclor, ampicillin-sulbactam, ertapenem, and piperacillin-tazobactam were the most commonly used antibiotics by patients preceding the onset of diarrhea. Among the patients who were hospitalized in an intensive care unit for more than 7 days, 83.3% were positive for CDI by PCR screening. If the PCR test is accepted as the reference: C. difficile Toxin A/B ELISA sensitivity and specificity were 67% and 94%, respectively, and GDH sensitivity and specificity were 100% and 75%, respectively.
  
  
  CONCLUSIONS
  Tests targeting C. difficile toxins are frequently applied for the purpose of diagnosing CDI in a clinical setting. However, changes in the temperature and reductant composition of the feces may affect toxin stability, potentially yielding false-negative test results. Therefore, employment of a GDH EIA, which has high sensitivity, as a screening test for the detection of toxigenic strains, may prevent false-negative results, and its adoption as part of a multistep diagnostic algorithm may increase accuracy in the diagnosis of CDIs.
authors.affiliation: Clinical Microbiology Laboratory, Türkan Özilhan Bornova State Hospital, İzmir, Turkey.
authors.forename: Özlem
authors.initials: Ö
authors.lastname: Yoldaş
authors.affiliation: Department of Medical Microbiology, Sakarya University Faculty of Medicine, Sakarya, Turkey.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Altındiş
authors.affiliation: Department of Medical Microbiology, Afyon Kocatepe University Faculty of Medicine, Afyon, Turkey.
authors.forename: Davut
authors.initials: D
authors.lastname: Cufalı
authors.affiliation: Department of Medical Microbiology, Afyon Kocatepe University Faculty of Medicine, Afyon, Turkey.
authors.forename: Gülşah
authors.initials: G
authors.lastname: Aşık
authors.affiliation: Department of Medical Microbiology, Afyon Kocatepe University Faculty of Medicine, Afyon, Turkey.
authors.forename: Recep
authors.initials: R
authors.lastname: Keşli
keywords: Clostridium difficile; PCR assay (GeneXpert®); antibiotic-associated diarrhea; glutamate dehydrogenase; toxin A/B