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A colorimetric microplate assay for determination of Staphylococcus aureus DNA is described. Linear padlock probes were designed to recognize target sequences. After DNA binding, the linear padlock probes were circularized by ligation and then hybridize with biotin-labeled capture probes. Biotin-labeled capture probes act as primers to initiate the RCA. The biotin-labeled RCA products hybridize with digoxin-labeled signal probes fixed on streptavidin-functionalized wells of a 96-well plate. To enhance sensitivity, an AuNP-anti-digoxigenin-POx-HRP conjugate was added to the wells and then bound to digoxin-labeled signalling probes. The oxidation of tetramethylbenzidine (TMB) by H2O2 produces a color change from colorless to blue via HRP catalysis. After the reaction was terminated, absorbance is measured at 450 nm. For target sequences of Staphylococcus aureus, the detection limit is 1.2 pM. For genomic DNA, the detection limit is 7.4 pg.μL−1. The potential application of the method was verified by analyzing spiked food samples. [ABSTRACT FROM AUTHOR]

The authors describe two kinds of immunochromatographic assays. The first is based on the use of dyed polymer microspheres (MICA), the another on the use of quantum dots (QICA). Both enable visual detection of enrofloxacin (ENR) in animal tissue and milk. Both the MICA and the QICA have visual limits of detection of 1 μg·L when working in buffer, of 5 μg·kg in case of animal tissue, and of 10 μg·L in case of milk. Other quinolones do not interfere. The MICA and QICA described here are convenient and fairly rapid in that the detection process (including sample pretreatment and assay) takes 20 min only which is far less than the commercial enzyme-linked immunosorbent assay (ELISA) test kit which requires 120 min. The MICA and QICA are more sensitive than the immunochromatographic assay using colloidal gold labels and the same polyclonal antibody. The results of analysis of spiked samples via MICA and QICA are in good agreement with those obtained by the commercial ELISA test kit. [ABSTRACT FROM AUTHOR]

A fluorometric aptamer-based method is described for the determination of aflatoxin B1 (AFB1). The fluorescent dye thioflavin T (ThT) forms a complex with the aptamer against AFB1 (aptamer/ThT), and the fluorescence of the complex is strongly enhanced. On addition of AFB1, it will bind to the aptamer and release ThT. The fluorescence of free ThT is much weaker. The fluorescence of the system, best measured at excitation/emission wavelengths of 440/487 nm, drops gradually in the AFB1 concentration range from 0.2 to 200 ng·mL−1, exhibiting good linearity. The detection limits are 0.2 ng·mL−1 in buffer solution, and 1 ng·mL−1 when applied to plant-derived food. The recovery of AFB1 from spiked foodstuff ranges from 74.7% to 121%. The assay can be performed within 20 min. Graphical abstract Schematic presentation of label-free thioflavin T (ThT)-based fluorescence aptasensor using aflatoxin B1(AFB1) aptamer/ThT G-quadruplex complex and the AFB1 aptamer/AFB1 complex fluorescent signal system for the rapid and sensitive detection of AFB1 in soy sauce, spirits, rice, corn and peanuts. [ABSTRACT FROM AUTHOR]

It is found that the fluorescence of carbon dots (CD) with an emission peak at 459 nm is strongly quenched by silver nanoparticles (AgNPs) with their absorption peak at 430 nm. The finding was applied in a fluorescence quenchometric lateral flow immunochromatographic assay for detection of zearalenone (ZEN) with CDs conjugated to ovalbumin (OVA) as donor signal probe and AgNP-Ab as acceptor signal probe. The assay has an LOD of 0.1 μg·L−1 for ZEN. This is 10 times better than the respective “turn-off” AgNP-based LFIA. In case of cereal samples and their products, the LODs range from 1 to 2.5 μg·kg−1. Only minor cross reactivity is found for fusarium toxins, and no cross-sensitivity for aflatoxin B1, T-2 mycotoxin, ochratoxin A, deoxynivalenol, and fumonisin B1. The assay represents a simple, sensitive, and rapid tool for determination of ZEN in cereal samples and their products.Schematic presentation of fluorescence quenching lateral flow immunochromatographic assay (FLFIA) based on carbon dots (CD) and silver nanoparticle (AgNP) fluorescence resonance energy transfer (FRET) system for the rapid high sensitive detection of zearalenone (ZEN) in cereal samples. [ABSTRACT FROM AUTHOR]