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Further characterization and independent validation of a DNA aptamer-quantum dot-based magnetic sandwich assay for Campylobacter

. 2017 Nov ; 62 (6) : 485-490. [epub] 20170324

Language English Country United States Media print-electronic

Document type Journal Article, Validation Study

Links

PubMed 28342148
DOI 10.1007/s12223-017-0520-0
PII: 10.1007/s12223-017-0520-0
Knihovny.cz E-resources

Previously reported DNA aptamers developed against surface proteins extracted from Campylobacter jejuni were further characterized by aptamer-based Western blotting and shown to bind epitopes on proteins weighing ~16 and 60 kD from reduced C. jejuni and Campylobacter coli lysates. Proteins of these approximate weights have also been identified in traditional antibody-based Western blots of Campylobacter spp. Specificity of the capture and reporter aptamers from the previous report was further validated by aptamer-based ELISA-like (ELASA) colorimetric microplate assay. Finally, the limit of detection of the previously reported plastic-adherent aptamer-magnetic bead and aptamer-quantum dot sandwich assay (PASA) was validated by an independent food safety testing laboratory to lie between 5 and 10 C. jejuni cells per milliliter in phosphate buffered saline and repeatedly frozen and thawed chicken rinsate. Such ultrasensitive and rapid (30 min) aptamer-based assays could provide alternative or additional screening tools to enhance food safety testing for Campylobacter and other foodborne pathogens.

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