Clostridium Difficile Toxin A and B ELISA Kit

Catalog No. ABIN1098188

Product Details

Target: Clostridium Difficile Toxin A and B

Reactivity: Clostridium difficile

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: ELISA for the simultaneous detection of Clostridium difficile toxin A and B in stool

Sample Type: Fecal

Analytical Method: Qualitative

Characteristics: The test is an in vitro diagnostic enzyme immunoassay for the detection of toxin A and toxin B produced by toxigenic strains of Clostridium difficile in human feces.

Components:

• 1x ELISA Plate 12x8 stripes
• 1 x 50 mL dilution buffer
• 1 x 2.0 mL Standard control Toxin A&B
• 1 x 7 mL conjugate
• 1 x 30 mL 10x washing buffer
• 1 x 14 mL substrate
• 1 x 7.5 stop solution with 0.5 M H2SO4
• 1x Manual

Material not included:

• Wash bottle
• Timer
• Paper towels or absorbent sheets
• Discard container
• Vortex mixer
• Microplate shaker
• Refrigerator
• Distilled water
• Microcentrifuge
• Microplate reader

Application Details

Application Notes:

• Sensitivity of the assay for TcdA: 0,5 ng/ml
• Sensitivity of the assay for TcdB: 1,0 ng/ml

Plate: Pre-coated

Sample Preparation:

Transfer about 50 µl liquid stool sample or take an equivalent amount (50 mg) of compact stool in 450 µl dilution buffer. Please only dilute the feces with the supplied dilution buffer, otherwise incorrect measurements can occur. Homogenize the suspension by suction and ejection from a disposable pipette or by vortexing. After leaving for a short time to allow sedimentation of stool particles the clarified supernatant can be used directly in the test. Automated equipment may be used with specimens that have been centrifuged 5 min by 2500 x g to remove any particulate matter.
Note: If overnight storage of the diluted sample is desirable, the storage should be done at -20 °C. Otherwise, in rare cases the test could lead to false positive results

Restrictions: For Research Use only

Handling

Preservative: ProClin

Handling Advice:

• Wear gloves for all manipulations with potentially contaminated or toxic suspensions.
• All reagents must be at room temperature prior to their use in the assay.

Storage: 4 °C

Storage Comment: Once opened, the kit is stable for at least 6 month when stored properly at 2-8 °C.

Expiry Date: 6 months

References

Hirano, Sakanaka, Yoshimi, Sugimoto, Eguchi, Yamauchi, Nara, Maeda, Ami, Gotoh, Katayama, Iida, Kato, Ohno, Fukiya, Yokota, Nishimoto, Kitaoka, Nakai, Kurihara: "Next-generation prebiotic promotes selective growth of bifidobacteria, suppressing Clostridioides difficile." in: Gut microbes, Vol. 13, Issue 1, pp. 1973835, (2022) (PubMed).

von Schwartzenberg, Bisanz, Lyalina, Spanogiannopoulos, Ang, Cai, Dickmann, Friedrich, Liu, Collins, Ingebrigtsen, Miller, Turnbaugh, Patterson, Pollard, Mai, Spranger, Turnbaugh: "Caloric restriction disrupts the microbiota and colonization resistance." in: Nature, Vol. 595, Issue 7866, pp. 272-277, (2021) (PubMed).

Edwards, Krall, McBride: "Strain-Dependent RstA Regulation of Clostridioides difficile Toxin Production and Sporulation." in: Journal of bacteriology, Vol. 202, Issue 2, (2020) (PubMed).

Dhillon, Johnson, Shannon, Greenwood, Roberts, Bustin: "Homogeneous and digital proximity ligation assays for the detection of Clostridium difficile toxins A and B." in: Biomolecular detection and quantification, Vol. 10, pp. 2-8, (2016) (PubMed).

Target Details

Target: Clostridium Difficile Toxin A and B

Background: C. difficile is an opportunistic anaerobic bacterium that grows in the intestine once the normal flora has been altered due to treatment with antibiotics. Subsequently, many patients develop gastrointestinal problems ranging from mild diarrhea to severe pseudomembranous colitis.

The clinical symptoms associated with the disease are believed to be primarily due to the tissue-damaging enterotoxin A (TcdA), whereas the cytotoxin (TcdB) is the one detected by the cell culture cytotoxicity assay.

Most strains produce both toxins, although clinically relevant toxin A negative/toxin B positive strains have been isolated with increasing frequency worldwide. Laboratory diagnosis of C. difficile infection is most commonly performed in a two-step algorithm: (1) screening of C. difficile presence using an immunoassay for the detection of C. difficile glutamate dehydrogenase (GDH) followed by (2) assaying the presence of toxins A/B using either an immunoassay and/or by PCR based techniques, the latter especially important in cases where the GDH test is positive but the toxin ELISA results negative. This could be the case for toxin production below the detection limit or capture of toxins by antitoxin antibodies.