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Xpert c difficile assay

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The Xpert C. difficile assay is a rapid molecular diagnostic test developed by Cepheid. It is designed to detect the presence of Clostridioides difficile (C. difficile) in patient samples. The assay utilizes real-time PCR technology to identify specific genetic markers associated with C. difficile infection.

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Lab products found in correlation

C diff quik chek complete, by Abbott (1 mentions) Vitek ms, by bioMérieux (1 mentions) Vitek 2, by bioMérieux (1 mentions) Genexpert system, by Cepheid (1 mentions) Max cdiff assay, by BD (1 mentions)

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Xpert C. difficile

14 protocols using xpert c difficile assay

1

Quantifying SCFA in C. difficile-positive Stools

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Human stool samples were collected from patients receiving care at Stanford Health Care between January 2015 and November 2015 and participating in an institutional review board (IRB)-exempt quality improvement project aimed at understanding the rates of C. difficile transmission in hematopoietic stem cell transplant patients. Samples either were from the patient’s first postadmission bowel movement or were collected at a frequency no more than once every 7 days postadmission. Samples were collected and immediately assayed for C. difficile TcdB using the Xpert C. difficile assay (Cepheid). Patients with unformed, C. difficile-positive stools were considered to have CDI. After this diagnostic procedure, residual deidentified samples (regardless of CDI status) were stored at 4°C for no more than 48 h and frozen at −80°C. Samples were subjected to targeted metabolomics, where the SCFAs acetate, propionate, and butyrate were quantified (see “SCFA quantification,” below).
Pensinger D.A., Fisher A.T., Dobrila H.A., Van Treuren W., Gardner J.O., Higginbottom S.K., Carter M.M., Schumann B., Bertozzi C.R., Anikst V., Martin C., Robilotti E.V., Chow J.M., Buck R.H., Tompkins L.S., Sonnenburg J.L, & Hryckowian A.J. (2023). Butyrate Differentiates Permissiveness to Clostridioides difficile Infection and Influences Growth of Diverse C. difficile Isolates. Infection and Immunity, 91(2), e00570-22.
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2

Rapid Identification of C. difficile

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Forty-nine stool samples were collected from CDI patients at the Poriya Baruch Padeh Medical Center, from November 2015 to May 2017. Specimens were analyzed by the Xpert C. difficile Assay (Cepheid, Sunnyvale, CA, USA), for rapid identification of Toxin B, Binary Toxin, and tcdC deletion presence.
An institutional review board, POR 0003-15, approved the study and informed consent was obtained from participants.
Mizrahi S., Hamo Z., Azrad M, & Peretz A. (2019). Molecular Characterization and Moxifloxacin Susceptibility of Clostridium difficile. Antibiotics, 8(3), 118.
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3

Cepheid Xpert C. difficile Assay Protocol

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Specimens were tested using the Cepheid Xpert C. difficile assay in accordance with the manufacturer's package insert. Briefly, wound cotton fiber swabs were inoculated with unpreserved fecal specimen and broken off into the Xpert elution reagent vial. The elution reagent vial was vortexed at high speed for 10 s and the entire volume subsequently transferred to the “S” chamber of the Xpert C. difficile assay cartridge using a transfer pipette. Results from the Xpert test were recorded for each specimen.
Mashock M.J., Faron M.L., Buchan B.W, & Ledeboer N.A. (2017). Evaluation of Copan FecalSwab as Specimen Type for Use in Xpert C. difficile Assay. Journal of Clinical Microbiology, 55(10), 3123-3129.
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4

Diarrheal Sample Analysis of CDI Patients

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The Institutional Review Board at the University of Virginia approved this study (IRB-HSR# 21646) with waiver of consent, as samples were de-identified remnants. Diarrheal samples from CDI+ (n = 20) and CDI− (n = 20) patients collected by the University of Virginia Medical Center between September 2019 and August 2020. CDI was diagnosed based on the presence of a conserved sequence of the tcdB gene, as determined by Xpert® C. difficile assay (Cepheid, Sunnyvale, CA, United States). To be included in the study, patients had to be at least 18 years old. Samples were stored in a −80°C freezer after collection until further processing.
Chen See J.R., Leister J., Wright J.R., Kruse P.I., Khedekar M.V., Besch C.E., Kumamoto C.A., Madden G.R., Stewart D.B, & Lamendella R. (2024). Clostridioides difficile infection is associated with differences in transcriptionally active microbial communities. Frontiers in Microbiology, 15, 1398018.
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5

Comparative CDI Testing Protocol

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The testing protocols for CDI were similar at both study sites. CDI cases were identified using a tiered approach in which clinical specimens submitted for C. difficile testing were first processed using a combined glutamate dehydrogenase (GDH) antigen enzyme immunoassay (EIA) and toxin A/B EIA (C. Diff Quik Chek Complete, Alere, Kansas City, MO). In instances of concordance (ie, both negative or both positive), the results are reported as negative or positive. In instances of discordance, reflex testing by polymerase chain reaction (PCR) for presence of toxin B gene (tcdB) using a commercial assay was conducted. The Xpert C. difficile assay (Cepheid, Sunnyvale, CA) was used at MGH. At UM, the GeneOhm assay (Becton Dickinson, Franklin Lakes, NJ) was used through 2013 and the Simplexa assay (Focus Diagnostics, Cypress, CA) was used thereafter. At MGH, formed stools were not rejected from the microbiology laboratory during the study period. At UM, formed stools were rejected beginning in May 2015. At each institution, the decision to test rested with the clinicians caring for the patient.
Oh J., Makar M., Fusco C., McCaffrey R., Rao K., Ryan E.E., Washer L., West L.R., Young V.B., Guttag J., Hooper D.C., Shenoy E.S, & Wiens J. (2018). A Generalizable, Data-Driven Approach to Predict Daily Risk of Clostridium difficile Infection at Two Large Academic Health Centers. Infection control and hospital epidemiology, 39(4), 425-433.
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6

Diagnosis of Clostridioides difficile Infection

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We diagnosed HO-CDI according to the United States Infectious Disease Society guidelines [13 (link)] when patients had associated symptoms (unformed stools ≥ three times/day) and a positive test for C. difficile (a two-step glutamate dehydrogenase assay and a NAAT or a NAAT alone) after 3 days of hospitalization. Patient stool samples were tested for the presence of the tcdB gene (which encodes toxin B) using the Xpert®C. difficile assay (Cepheid; Sunnyvale, CA, USA) according to the manufacturer’s instructions [20 (link)].
Yun J.H., Park G.E, & Ki H.K. (2021). Correlation between antibiotic consumption and the incidence of healthcare facility-onset Clostridioides difficile infection: a retrospective chart review and analysis. Antimicrobial Resistance and Infection Control, 10, 117.
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7

Xpert C. difficile Assay Evaluation

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The Xpert C. difficile assay (Cepheid Inc., Sunnyvale, CA) was performed using the Cepheid GeneXpert Systems. Sixty of the 96 GDH-positive CCNA-negative samples were tested with the Xpert C. difficile assay by the Leeds laboratory as part of an internal evaluation. At bioMérieux, the testing of the 36 GDH-positive CCNA-negative samples was completed, as well as additional testing for samples that gave discordant results between CCNA and the SIMOA assay.
Banz A., Lantz A., Riou B., Foussadier A., Miller M., Davies K, & Wilcox M. (2018). Sensitivity of Single-Molecule Array Assays for Detection of Clostridium difficile Toxins in Comparison to Conventional Laboratory Testing Algorithms. Journal of Clinical Microbiology, 56(8), e00452-18.
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8

Comparative Diagnostic Evaluation for CDI

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Three hundred diarrhoeal stool specimens submitted for CDI diagnosis have been included consecutively, if they were less than 24 h old and of sufficient volume to allow testing with all study-related assays in addition to routine testing. The latter comprised an initial screening by a GDH EIA (C. diff Chek-60; TechLab, Blacksburg, VA, USA). In case of a positive result confirmatory PCR testing (Xpert C. difficile assay; Cepheid, Sunnyvale, CA, USA) was conducted. Specimens were given a study protocol number by the routine staff and were, thereby anonymised, forwarded to the scientific staff for further analysis by TC, CCTA, EIA for toxins A and B and the two CLIAs, one for GDH and the other for toxins A and B. The results of routine testing were communicated to the clinical ward; those obtained by the study-related tests were not disclosed, and therefore, did not impact the patient’s management.
Makristathis A., Zeller I., Mitteregger D., Kundi M, & Hirschl A.M. (2017). Comprehensive evaluation of chemiluminescent immunoassays for the laboratory diagnosis of Clostridium difficile infection. European Journal of Clinical Microbiology & Infectious Diseases, 36(7), 1253-1259.
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9

Detection of Enteric Pathogens

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Bacterial culture (sheep blood agar, MacConkey agar, xylose lysine deoxycholate agar, and Campylobacter CVA agar) was used for detection of Salmonella, Shigella, and Campylobacter species following standard methods. The Cepheid Xpert C difficile assay was used for detection of toxigenic C difficile, and the Premier Adenoclone-Type 40/41 and Premier Rotaclone enzyme immunoassay test kits (Meridian Bioscience, Inc., Cincinnati, OH) were used for the detection of adenovirus serotypes 40 and 41 and rotavirus, respectively. For detection of Giardia lamblia, the ProSpecT Giardia Microtiter assay (Remel, Inc., Lenexa, KS) was used.
Bok K., Prevots D.R., Binder A.M., Parra G.I., Strollo S., Fahle G.A., Behrle-Yardley A., Johnson J.A., Levenson E.A., Sosnovtsev S.V., Holland S.M., Palmore T.N, & Green K.Y. (2016). Epidemiology of Norovirus Infection Among Immunocompromised Patients at a Tertiary Care Research Hospital, 2010–2013. Open Forum Infectious Diseases, 3(3), ofw169.
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10

Rapid Detection of Toxigenic C. difficile

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Samples were processed using a rapid detection kit for toxigenic C. difficile. This rapid test consists of glutamate dehydrogenase antigen and toxins A and B detection by immunochromatography (C Diff Quik-Chek Complete assay, TechLab, Blacksburg, VA, USA) (Sensitivity 90.5%, specificity 93.1%, predictive positive value 76.4%, predictive negative value 97.6% in comparison with bacterial culture) and a real-time polymerase chain reaction (PCR) assay of the C. difficile toxin B gene (Xpert.C. difficile Assay, GeneXpert, Cepheid, Sunnyvale, CA, USA) (Sensitivity 98.79%, specificity 90.82%, predictive positive value 56.58%, predictive negative value 99.83% in comparison with bacterial culture).
In addition, all samples were cultured on C. difficile selective agar (bioMeriéux, Marcy l’Etoile, France). Suspected toxigenic C. difficile colonies were confirmed using immunochromatography (C Diff Quik-Chek Complete assay, TechLab, Blacksburg, VA, USA).
Vázquez-Cuesta S., Lozano García N., Fernández A.I., Olmedo M., Kestler M., Alcalá L., Marín M., Bermejo J., Díaz F.F., Muñoz P., Bouza E, & Reigadas E. (2023). Microbiome profile and calprotectin levels as markers of risk of recurrent Clostridioides difficile infection. Frontiers in Cellular and Infection Microbiology, 13, 1237500.
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