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Pubmed ID :30158195
Publication Date : //

Ultrasensitive Detection of Toxins A and B Using Automated Single Molecule Counting Technology.


Current tests for the detection of (formerly ) free toxins in feces lack sensitivity, while nucleic acid amplification tests lack clinical specificity. We have evaluated the Singulex Clarity® C. diff toxins A/B assay (currently in development), an automated and rapid ultrasensitive immunoassay powered by Single Molecule Counting technology, for detection of toxins A (TcdA) and B (TcdB) in stool. Analytical sensitivity, analytical specificity, repeatability, and stability for the assay were determined. In a clinical evaluation, frozen stool samples from 311 patients with suspected CDI were tested with the Clarity C. diff toxins A/B assay with an established cutoff. Samples were tested with Xpert® , and PCR+ samples were tested with an enzyme immunoassay (EIA; C. Diff Quik Chek Complete®). EIA- samples were further tested with cell cytotoxicity neutralization assay. Limits of detection for TcdA and TcdB were 0.8 and 0.3 pg/mL in buffer, and 2.0 and 0.7 pg/mL in stool, respectively. The assay demonstrated reactivity to common strains, did not show cross-reactivity to common gastrointestinal pathogens, was robust against common interferents, allowed for detection in fresh and frozen stool samples and after three freeze-thaw cycles, and provided results with high reproducibility. Compared to multistep PCR-and-toxin testing, the Singulex Clarity C. diff toxins A/B assay yielded 97.7% sensitivity and 100% specificity. The Singulex Clarity C. diff toxins A/B assay is ultrasensitive and highly specific and may offer a standalone solution for rapid detection and quantitation of free toxins in stool.

Authors : Sandlund Johanna , Bartolome Amelita , Almazan Anna , Tam Stanley , Biscocho Sheryl , Abusali Salina , Bishop Jeff , Nolan Niamh , Estis Joel , Todd John , Young Stephen , Senchyna Fiona , Banaei Niaz ,

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