Alternative Causes of Infectious Diarrhea in Patients with Negative Tests for Clostridoides Difficile

Background Hospitalized patients with diarrhea who have a negative Clostridoides difficile (C. difficile) test are not routinely evaluated for alternative causes of infectious diarrhea. This study assessed for potential infectious causes of diarrhea in hospitalized patients with an order for repeat C. difficile toxin enzyme immunoassay (tEIA) testing after an initial tEIA test was negative. Methods For patients age ≥18 years who had a second C. difficile tEIA test ordered within 96 h after a negative tEIA test, remnant fecal specimens from the first (negative) tEIA test were evaluated using the BioFire FilmArray Gastrointestinal Panel PCR, C. difficile toxigenic culture, and culture on a blood agar plate (BAP) to identify other potential causes of infectious diarrhea. Growth of organisms on the BAP was also used to assess potential disruptions in the gastrointestinal microbiota. Results Among 84 remnant specimens, toxigenic C. difficile was identified in 9 (11%) by culture or PCR, while potential alternative causes of infectious diarrhea, including norovirus, rotavirus, enteropathogenic Escherichia coli, and Salmonella, were identified in 11 specimens (13%) by PCR. For the majority of patients, no infectious cause of diarrhea was identified, but 84% exhibited disrupted gastrointestinal microbiota, which may contribute to diarrhea. Conclusions When a hospitalized patient has a negative C. difficile tEIA test but continues to have diarrhea, alternative infectious and noninfectious causes of diarrhea should be considered. If the patient has clinical signs and symptoms suggestive of infection or risk factors for gastrointestinal infection, laboratory testing for other etiologic agents may be appropriate.

Significance of a polymerase chain reaction method in the detection of Clostridioides difficile

Objectives. Clostridioides difficile (CD) is the most common cause of nosocomial diarrhea. Detection of CD toxin in patients’ faecal samples is the traditional rapid method for the diagnosis of CD infection. Various testing algorithms have been proposed: an initial screening test using a rapid test, and a confirmatory test (cytotoxicity neutralization assay, toxigenic culture, nucleic acid amplification test) for discordant results. The aim of this study was to evaluate the effectiveness of a two-step algorithm using an immunochromatographic test followed of a polymerase chain reaction (PCR). Material and methods. The specimens have been tested according to the following schedule: 1) Step one: All samples were tested for detection of glutamate dehydrogenase antigen (GDH) and toxin A/B using the C. diff QUIK CHEK Complete test. All GDH and toxins positive results were considered CD positives; 2) Step two: When the results were discrepant (only GDH+ or toxins+), the samples were confirmed using the PCR test BD MAX Cdiff. All PCR positive results were considered CD positives. Results. A total of 2,138 specimens were initially tested. 139 were positive for GDH and toxins. 160 discrepant results (148 GDH+ and 12 toxins+) were tested by PCR, 117 were positive (107/148 GDH+ and 10/12 toxins+). Conclusions. The implementation of a PCR method showed an increase de 117 positive results (73.1% of discrepant). Considering the sensitivity of C.diff QUIK CHEK (instructions of manufacturer), the GDH discrepant results may be false negatives, y the samples PCR and toxins positives may be real positives results.

Latent class analysis for the diagnosis of Clostridioides difficile infection

Background Clostridioides difficile infection (CDI) is an opportunistic disease that lacks a gold standard test. Nucleic acid amplification tests (NAATs) such as real-time PCR demonstrate excellent an limit of detection (LOD) whereas antigenic methods are able to detect free toxin. Latent class analysis (LCA) provides an unbiased statistical approach to resolving true disease. Methods A cross-sectional study was conducted with suspected CDI patients (n=96). Four commercial real-time PCR tests, toxin antigen detection by enzyme immunoassay (EIA), toxigenic culture, and fecal calprotectin were performed. CDI clinical diagnosis was determined by consensus majority of three experts. LCA was performed using laboratory and clinical variables independent of any gold standard. Results Six LCA models were generated to determine CDI probability using four variables including toxin EIA, toxigenic culture, clinical diagnosis, and fecal calprotectin levels. Three defined zones as a function of real-time PCR cycle threshold (Ct) were identified using LCA: CDI likely (>90% probability), equivocal (<90% and >10%), CDI unlikely (<10%). A single model comprising toxigenic culture, clinical diagnosis, and toxin EIA showed the best fitness. The following Ct cut-offs for four commercial test platforms were obtained using this model to delineate three CDI probability zones: [GeneXpert ® : 24.00, 33.61], [Simplexa ® 28.97, 36.85], [Elite MGB ® 30.18, 37.43], and [BD Max ™ 27.60, 34.26]. Conclusion The clinical implication of applying LCA to CDI is to report Ct values assigned to probability zones based on the commercial real-time PCR platform. A broad range of equivocation suggests clinical judgement is essential to the confirmation of CDI.

Association Between Federal Value-Based Incentive Program Implementation and Hospital-Onset C. difficile Infection Rates

Background: The Hospital-Acquired Conditions Reduction Program (HACRP) and Hospital Value-Based Purchasing (HVBP) are federal value-based incentive programs that financially reward or penalize hospitals based on quality metrics. Hospital-onset C. difficile infection (HO-CDI) rates reported to the CDC NHSN became a target quality metric for both HACRP and HVBP in October 2016, but the impact of these programs on HO-CDI rates is unknown. Methods: We used an interrupted time-series design to examine the association between HACRP/HVBP implementation in October 2016 and quarterly rates of HO-CDI per 10,000 patient days among incentive-eligible acute-care hospitals conducting facility-wide HO-CDI NHSN surveillance between January 2013 and March 2019. Generalized estimating equations were used to fit negative binomial regression models to assess for immediate program impact (ie, level change) and changes in the slope of HO-CDI rates, controlling for each hospital’s predominant method for CDI testing (nucleic acid amplification including PCR (NAAT), enzyme immunoassay for toxin (EIA), or other testing method including cell cytotoxicity neutralization assay and toxigenic culture). Results: Of the 265 study hospitals studied, most were medium-sized (100–399 beds, 55%), not-for-profit (77%), teaching hospitals (70%), and were located in a metropolitan area (87%). Compared to EIA, rates of HO-CDI were higher when detected by NAAT (incidence rate ratio [IRR], 1.55; 95% CI, 1.41–1.70) or other testing methods (IRR, 1.47; 95% CI, 1.26–1.71). Controlling for CDI testing methods, HACRP/HVBP implementation was associated with an immediate 6% decline in HO-CDI rates (IRR, 0.94; 95% CI, 0.89–0.99) and a 4% decline in slope per year-quarter thereafter (IRR, 0.96; 95% CI, 0.95–0.97) (Fig. 1). Conclusions: HACRP/HVBP implementation was associated with both immediate and gradual improvements in HO-CDI rates, independent of CDI testing methods of differing sensitivity. Future research may evaluate the precise mechanisms underlying this improvement and if this impact is sustained in the long term.Funding: NoneDisclosures: None

Comparative Study of Clostridium difficile Clinical Detection Methods in Patients with Diarrhoea

Objectives. The aim of this study was to evaluate the clinical application of three methods for detecting Clostridium difficile in fecal samples. Methods. One hundred and fifty fecal specimens were collected and tested for C. difficile using three methods: (1) the toxigenic culture (TC); (2) the VIDAS enzyme immunoassay (EIA): the VIDAS glutamate dehydrogenase (GDH) assay and toxin A/B assay were used to detect GDH antigen and A/B toxin; and (3) the GeneXpert PCR assay. The toxigenic culture was used as a reference to evaluate the performance of the VIDAS EIA and the GeneXpert PCR assay. Results. Of 150 specimens, 26 carried both A and B toxin genes, and none of the samples were positive for the binary toxin gene. Toxin-producing C. difficile was found in 17.3% (26/150) of the samples. Thirty-seven GDH-positive samples were detected using the VIDAS GDH assay, and 15 toxin-positive samples were detected using the VIDAS toxin A/B assay. The GeneXpert PCR assay was used to detect C. difficile in 79 specimens simultaneously, and a total of 18 positive specimens were detected. Conclusion. The VIDAS GDH assay is useful for initial screening of C. difficile. The GeneXpert PCR assay is a simple and quick method.

Evaluation of a Gastrointestinal Pathogen Panel Immunoassay in Stool Testing of Patients with Suspected Clostridioides (Clostridium) difficile Infection

ABSTRACT Clostridioides (Clostridium) difficile infection (CDI) is the most common causative pathogen of health care-associated gastrointestinal infections; however, due to the overlap of clinical symptoms with those of other causes of acute gastroenteritis, the selection of the most appropriate laboratory test is difficult. From April to October 2018, 640 stool samples requested for CDI testing were examined using the mariPOC CDI and Gastro test (ArcDia), which allows the detection of C. difficile glutamate dehydrogenase (GDH) and toxin A/B, norovirus genogroups GI and GII.4, rotavirus, adenovirus, and Campylobacter spp. In parallel, the C. Diff Quik Chek Complete test (Alere) was used as a routine diagnostic assay, and C. difficile toxigenic culture was used as a reference method. The sensitivity of the mariPOC CDI and Gastro test was comparable to that of C. Diff Quik Chek Complete for the detection of GDH (96.40% [95% confidence interval {CI}, 91.81% to 98.82%] versus 95.68% [95% CI, 90.84 to 98.40%]; P = 1.00) and was higher for the detection of toxin A/B (66.67% [95% CI, 57.36 to 75.11%] versus 55.56% [95% CI, 46.08 to 64.74%]; P = 0.00). The specificity of the mariPOC CDI and Gastro test was lower than that of C. Diff Quik Chek Complete for GDH detection (95.21% [95% CI, 92.96% to 96.91%] versus 97.60% [95% CI, 95.85% to 98.76%]; P = 0.04) and comparable to that of C. Diff Quik Chek Complete for toxin A/B detection (99.24 [95% CI, 98.05% to 99.79%] versus 99.81% [95% CI, 98.94% to 100.0%]; P = 0.37). In 29 cases (4.53%), other causative agents of diarrhea were detected (Campylobacter spp. [n = 17], rotavirus [n = 7], and norovirus genogroup GII.4 [n = 5]).

Influence of Diagnostic Method on Outcomes in Phase 3 Clinical Trials of Bezlotoxumab for the Prevention of Recurrent Clostridioides difficile Infection: A Post Hoc Analysis of MODIFY I/II

Background The optimum diagnostic test method for Clostridioides difficile infection (CDI) remains controversial due to variation in accuracy in identifying true CDI. This post hoc analysis examined the impact of CDI diagnostic testing methodology on efficacy outcomes in phase 3 MODIFY I/II trials. Methods In MODIFY I/II (NCT01241552/NCT01513239), participants received bezlotoxumab (10 mg/kg) or placebo during anti-CDI treatment for primary/recurrent CDI (rCDI). Using MODIFY I/II pooled data, initial clinical cure (ICC) and rCDI were assessed in participants diagnosed at baseline using direct detection methods (enzyme immunoassay [EIA]/cell cytotoxicity assay [CCA]) or indirect methods to determine toxin-producing ability (toxin gene polymerase chain reaction [tgPCR]/toxigenic culture). Results Of 1554 participants who received bezlotoxumab or placebo in MODIFY I/II, 781 (50.3%) and 773 (49.7%) were diagnosed by tgPCR/toxigenic culture and toxin EIA/CCA, respectively. Participants diagnosed by toxin EIA/CCA were more likely to be inpatients, older, and have severe CDI. In bezlotoxumab recipients, ICC rates were slightly higher in the toxin EIA/CCA subgroup (81.7%) vs tgPCR/toxigenic culture (78.4%). Bezlotoxumab significantly reduced the rCDI rate vs placebo in both subgroups; however, the magnitude of reduction was substantially larger in participants diagnosed by toxin EIA/CCA (relative difference, –46.6%) vs tgPCR/toxigenic culture (–29.1%). In bezlotoxumab recipients, the rCDI rate was lower in the toxin EIA/CCA subgroup (17.6%) vs tgPCR/toxigenic culture (23.6%; absolute difference, –6.0%; 95% confidence interval, –12.4 to 0.3; relative difference, –25.4%). Conclusions Diagnostic tests that detect fecal C. difficile toxins are of fundamental importance to accurately diagnosing CDI, including in clinical trial design, ensuring that therapeutic efficacy is not underestimated.

Evaluation of Risk Factors forClostridium difficileInfection Based on Immunochromatography Testing and Toxigenic Culture Assay

ABSTRACTIn recent years, the diagnostic method of choice forClostridium difficileinfection (CDI) is a rapid enzyme immunoassay in which glutamate dehydrogenase (GDH) antigen andC. difficiletoxin can be detected (C. diffQuik Chek Complete; Alere Inc.) (Quik Chek). However, the clinical significance remains unclear in cases that demonstrate a positive result for GDH antigen and are negative for toxin. In this study, we used the Quik Chek test kit on fecal samples, with an additional toxin detection step using a toxigenic culture assay for the aforementioned cases. CDI risk factors were assessed among the 3 groups divided by the Quik Chek test results. The study involved 1,565 fecal samples from patients suspected to have CDI who were hospitalized during the period of April 2012 to March 2014. The 3 groups were defined as follows: both GDH antigen positive and toxin positive (by Quik Chek test) (toxin-positive [TP] group,n= 109), both GDH antigen and toxin negative (toxin-negative [TN] group,n= 111), and positive only for GDH antigen but toxin positive with subsequent toxigenic culture (toxigenic culture [TC] group,n= 72). The gender, age, number of hospitalization days, white blood cell (WBC) counts, serum albumin levels, body mass index (BMI), fecal consistency, and use of antibacterials and proton pump inhibiters (PPIs) were analyzed. The positive rate for the fecal direct Quik Chek test was 7.0% (109/1,565 cases). However, toxigenic culture assays using the Quik Chek test for only the GDH-antigen-positive/toxin-negative samples were 35.3% positive (72/204 cases). As a result, the true positive rate forC. difficiletoxin detection was estimated to be 11.6% (181/1,565 cases). Moreover, significant differences (P< 0.05) in the number of hospitalization days (>50 days), WBC counts (>10,000 WBCs/μl), and use of PPIs comparing the TN, TP, and TC groups, were observed. The odds ratios (ORs) for the development of CDI were 1.61 (95% confidence interval [CI], 0.94 to 2.74) and 2.98 (95% CI, 1.59 to 5.58) for numbers of hospitalization days, 2.16 (95% CI, 1.24 to 3.75) and 2.24 (95% CI, 1.21 to 4.14) for WBC counts, and 9.03 (95% CI, 4.9 to 16.6) and 9.15 (95% CI, 4.59 to 18.2) for use of PPIs in the TP and TC groups, respectively. These findings demonstrated that the use of PPIs was a significant risk factor for CDI development. Moreover, antibacterials such as carbapenems, cephalosporins, and fluoroquinolones were demonstrated to be risk factors. In conclusion, identification of the TC group of patients is thought to be important, as this study demonstrates that this group bears the same high risk of developing CDI as the TP group.