Indirect Competitive Enzyme-Linked Immunosorbent Assay Based on Broad-Spectrum Antibody for Simultaneous Determination of Thirteen Fluoroquinolone Antibiotics in Rana catesbeianus.

Fluoroquinolone (FQ) is a type of widely used antibiotic in agriculture and aquaculture, and exposure to low doses of FQs may result in the transfer of resistance between animal and human pathogens. Based on the optimization of the operating parameters, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) standard curve was constructed for the simultaneous detection of 13 FQs, including enrofloxacin (ENR), ciprofloxacin (CIP), sarafloxacin (SAR), ofloxacin (OFL), norfloxacin (NOR), pefloxacin mesylate (PM), pefloxacin (PEF), enoxacin (ENX), marbofloxacin (MAR), fleroxacin (FLE), lomefloxacin (LOM), danofloxacin (DAN), and difloxacin (DIF). The limit of detection (LOD, computed as IC10) and sensitivity (IC50) of the ic-ELISA for ENR were 0.59 µg/L and 19.23 µg/L, respectively. The precision and dependability of the detection results of this ic-ELISA were properly verified by HPLC in Rana catesbeianus samples. This indicated that the established ic-ELISA approach could be utilized to determine the FQs in Rana catesbeianus. In addition, this ic-ELISA, based on a broad-spectrum antibody, provides a technical reference and potential strategy for an immunoassay of hazard factors with similar structure.

A self-designed device integrated with a Fermat spiral microfluidic chip for ratiometric and automated point-of-care testing of anthrax biomarker in real samples.

A desirable lanthanide-based ratiometric fluorescent probe was designed and integrated into a self-designed Fermat spiral microfluidic chip (FS-MC) for the automated determination of a unique bacterial endospore biomarker, dipicolinic acid (DPA), with high selectivity and sensitivity. Here, a blue emission wavelength at 425 nm was generated in the Fermat spiral structure by mixing the europium (Eu3+) and luminol to form the Eu3+/Luminol sensing probe. DPA in the reservoir can be used to specifically bind to Eu3+ under the negative pressure and transfer energy from DPA to Eu3+ sequentially via an antenna effect, thus resulting in a significant increase in the red fluorescence emission peak at 615 nm. According to the fluorescence intensity ratio (F615/F425), a good linearity can be obtained with increasing the concentration of DPA from 0 to 200 µM with a limit of detection as low as 10.11 nM. Interestingly, the designed FS-MC can achieve rapid detection of DPA in only 1 min, reducing detection time and improving sensitivity. Furthermore, a self-designed device integrated with the FS-MC and a smartphone color picker APP was employed for the rapid automatic point-of-care testing (POCT) of DPA in the field, simplifying complex processes and reducing testing times, thus confirming the great promise of this ready-to-use measurement platform for in situ inspection.

A Smartphone Colorimetric Sensor Based on Pt@Au Nanozyme for Visual and Quantitative Detection of Omethoate.

A smartphone colorimetric sensor based on the Pt@Au nanozyme was successfully developed for the visual and quantitative detection of omethoate in fruit and vegetables. The anti-omethoate antibody was conjugated on the surface of the Pt@Au nanozyme as a catalytic functional signal probe, and coating antigen conjugated on the surface of magnetic polystyrene microspheres (MPMs) was used as a separation capture probe. In the sensing system, when the catalytic functional signal probe was combined with a separation capture probe containing no omethoate, the visible blue color appeared with the addition of tetramethylbenzidine (TMB) chromogenic solution, and the maximum B value of the sensing system was obtained via the smartphone. With increasing concentrations of omethoate, the visualization of the sensing system decreased, and the B-value obtained via the smartphone dropped. Under optimal detection conditions, the omethoate could be detected in a linear range of 0.5-50 µg/L (R2 = 0.9965), with a detection limit of 0.01 µg/L. The accuracy and reliability of the detection results of this colorimetric sensor were successfully confirmed by enzyme linked immunosorbent assay (ELISA) and gas chromatography. This colorimetric sensor provides a technical reference and potential strategy for the immunoassay of hazard factors in resource-scarce laboratories.

Determination Methods of the Risk Factors in Food Based on Nanozymes: A Review.

Food safety issues caused by foodborne pathogens, chemical pollutants, and heavy metals have aroused widespread concern because they are closely related to human health. Nanozyme-based biosensors have excellent characteristics such as high sensitivity, selectivity, and cost-effectiveness and have been used to detect the risk factors in foods. In this work, the common detection methods for pathogenic microorganisms, toxins, heavy metals, pesticide residues, veterinary drugs, and illegal additives are firstly reviewed. Then, the principles and applications of immunosensors based on various nanozymes are reviewed and explained. Applying nanozymes to the detection of pathogenic bacteria holds great potential for real-time evaluation and detection protocols for food risk factors.