Aptamer–Magnetic Bead Quantum Dot Sandwich Assays for Foodborne Pathogen Detection: Pros, Cons, and Lessons Learned

Abstract

DNA and RNA aptamers have been extensively investigated as potential competitors for antibodies for a variety of applications including food safety testing. Ultrasensitive fluorescence detection of foodborne pathogenic bacteria as low as 1-10 cells/mL has been achieved using aptamers coupled to quantum dots in clear pristine buffers for environmental sample detection. Quantum dots offer other advantages, including single UV or blue light source multiplex (multicolored) detection. However, quantum dots can exhibit decreased fluorescence in some food matrixes and even completely fail to fluoresce in some fatty matrixes, as documented in this report. Given the need to detect substances in complex food matrixes (and from data reported elsewhere), aptamer-magnetic bead pull-down methods followed by enzymatic/fluorometric- or PCR-based detection methods may be more robust methods for testing in foods or enrichment cultures. Other lessons learned, including the initial choice of aptamer targets to enhance assay specificity, are also discussed.