The Xpert C. difficile/Epi assay (Cepheid, CA) was performed as described by the manufacturer. Briefly, each stool sample positive for culture was transferred into a sample reagent vial. The vial was vortexed for 10 s, and the solution was pipetted into the cartridge by using a Pasteur pipette. The cartridge was placed on the Xpert instrument, and the assay was performed using the GeneXpert C. difficile assay program.
Xpert c difficile epi assay
Manufactured by Cepheid
Sourced in United States
The Xpert C. difficile/Epi assay is a rapid, automated molecular diagnostic test that detects the presence of Clostridioides difficile (C. difficile) and the binary toxin gene in patient samples. The assay utilizes real-time PCR technology to provide accurate and timely results, aiding in the diagnosis of C. difficile infection.
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4 protocols using xpert c difficile epi assay
1
Xpert C. difficile/Epi Assay Protocol
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Clostridium difficile Infection Diagnosis in Slovakia
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The six-month study was carried out from 1 January to 30 June 2016 in a tertiary-care center in the northern part of Slovakia. The hospital had 726 beds and 12,575 admissions during the study period.
Unformed stool samples from patients with a suspected CDI were tested for the presence of toxins A/B (CerTest Clostridium difficile GDH + Toxin A + B one-step combo card test, Zaragoza Spain). The toxin A/B-positive stool samples were investigated using the Xpert®C. difficile/Epi assay (Cepheid, Sunnyvale, CA, USA) that detects the presence of tcdB (toxin B) and cdtB (binary toxin) genes and the deletion at position 117 in the tcdC gene (glycosylating toxin anti-sigma factor). All toxin A/B-positive stool samples were simultaneously cultured anaerobically on selective media for C. difficile (Brazier, Hampshire, Oxoid, UK).
Unformed stool samples from patients with a suspected CDI were tested for the presence of toxins A/B (CerTest Clostridium difficile GDH + Toxin A + B one-step combo card test, Zaragoza Spain). The toxin A/B-positive stool samples were investigated using the Xpert®C. difficile/Epi assay (Cepheid, Sunnyvale, CA, USA) that detects the presence of tcdB (toxin B) and cdtB (binary toxin) genes and the deletion at position 117 in the tcdC gene (glycosylating toxin anti-sigma factor). All toxin A/B-positive stool samples were simultaneously cultured anaerobically on selective media for C. difficile (Brazier, Hampshire, Oxoid, UK).
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Diagnosis of Clostridioides difficile Infection
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Microbiological diagnosis of C. difficile disease was performed by the use of enzyme immunoassays combining detection of C. difficile glutamate dehydrogenase (GDH) and toxin A/B antigens in stool specimens, with the commercial methods TechLab C. Diff Quik Chek Complete or Meridian Bioscience Immunocard C. difficile GDH and Vidas C. difficile Toxin A/B, according to the testing algorithms established at the individual study sites. All stool specimens were also tested with the Cepheid Xpert C. difficile/Epi assay, which is a multiplex realtime PCR that detects tcdB, the binary toxin gene (cdt), and the tcdC gene deletion at nucleotide 117, to identify the PCRribotype 027 strain, also called the 027/NAP1/BI strain [8] .
Blood specimens were processed with the automated BACTEC system (Becton Dickinson Diagnostic Instruments, Sparks, MD, USA).
Blood specimens were processed with the automated BACTEC system (Becton Dickinson Diagnostic Instruments, Sparks, MD, USA).
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Rapid Diagnosis of C. difficile Infection
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Clinical diagnosis of CDI was suspected when patients were hospitalized for more than 48 h and had >3 loose stools in the previous 24 h. In the Hospital Civil of Guadalajara, CDI was confirmed by real-time PCR using the Xpert ® C. difficile/Epi assay (Cepheid, Sunny Vale, CA, USA) and in the Hospital Universitario, the diagnosis was confirmed by the use of the Meridian ImmunoCard ® Toxins A&B (Meridian Bioscience, Inc., Memphis, TN, USA). Some patients were additionally diagnosed by the Xpert ® C. difficile/Epi assay in Hospital Universitario only at physicians' request.
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