Dramatic fluorescence enhancement of PCN-224 and its application in "turn off" immunoassay for sensitive detection of E. coli O157:H7 in milk.

In this study, a type of luminescent porous coordination network-224 (PCN-224) in alkaline conditions was synthesized with the dramatic fluorescence enhancement by 20.4 times, which was explained by the fact that the decrease of Zr4+ content in alkaline conditions resulted in the partial recovery of the electron cloud density of 4,4',4'',4'''-(Porphine-5,10,15,20-tetrayl) tetrakis(benzoic acid) (TCPP). Given the large overlap between the excitation spectrum of PCN-224 and the absorption band of Ag nanoparticles (Ag NPs), the coating of the Ag layer on PCN-224 triggered the fluorescence quenching effect, which was applied to "turn off" fluorescence immunoassay for sensitive detection of Escherichia coli O157:H7 (E. coli O157:H7) in milk. The proposed immunoassay reached a low limit of detection (LOD) of 3.3 × 102 CFU mL-1, 29.7 times more sensitive than the conventional ELISA. It will provide a novel alternative strategy for sensitively detecting pathogenic bacteria in the field of food safety.

Lateral flow immunoassay based on polydopamine-coated metal-organic framework for the visual detection of enrofloxacin in milk.

Herein, we report a new polydopamine (PDA)-coated metal-organic framework (MOF) as a label to improve the sensitivity of lateral flow immunoassay (LFIA). The MOF, UiO-66-NH2, was synthesized via the hydrothermal method, and it exhibited the advantageous features of ordered pore structure, strong absorbance, and high specific surface area. Subsequently, UiO-66-NH2 was coated with PDA to improve the antibody coupling effectivity and light absorption ability. The optical intensity and antibody coupling efficiency of UiO-66@PDA were superior to those of gold nanoparticles (AuNPs). Under the optimum condition, the limit of detection and cutoff value of UiO-66@PDA-LFIA in detecting enrofloxacin were 0.045 and 1.0 ng/mL, respectively, which were lower than those of AuNPs-LFIA (0.095 and 5 ng/mL). The recoveries of UiO-66@PDA-LFIA in low-fat milk and whole milk were 85.6-107.4% and 79.3-115.5%, respectively, with coefficients of variation of 2.91-9.59% and 3.91-11.8%, respectively, as further confirmed by liquid chromatography-tandem mass spectrometry. These results indicate that UiO-66@PDA can be used as a novel probe for LFIA development and applications. Graphical abstract.

The relationship between serum uric acid levels and glomerular ischemic lesions in patients with Immunoglobin A nephropathy-a analytical cross-sectional study.

BACKGROUND: To investigate the relationship between serum uric acid levels and glomerular ischemic lesions in patients with immunoglobulin A nephropathy (IgAN) and the relevant risk factors. METHODS: A total of 86 patients with IgAN and normal renal functions were divided into a hyperuricemia group and a normal serum uric acid group (control group). These patients were further divided into a glomerular ischemic lesions group and a non-glomerular ischemic lesions group (control group) based on the renal biopsy results. The relationship between serum uric acid levels and glomerular ischemic lesions was analysed. RESULTS: In patients with IgAN, the prevalence or occurrence of glomerular ischemic lesions was significantly higher in the hyperuricemia group compared with the normal serum uric acid group. Elevated serum uric acid levels are independently associated with glomerular ischemic disease. CONCLUSION: Hyperuricemia in patients with IgAN may lead to glomerular ischemic lesions, and lowering serum uric acid levels may delay the progression of IgAN.

A novel method based on Ag-Au nanorings with tunable plasmonic properties for the sensitive detection of amantadine.

To prevent the toxic effect of amantadine (AMD) on humans, a sensitive and direct detection method is required. The conventional enzyme-linked immunosorbent assay (ELISA) usually shows a monochromatic gradient color variation with the concentration of the target; hence, it is not a sensitive method for naked-eye detection. In this work, Ag-Au nanorings with highly tunable plasmon properties were synthesized as colorimetric nanosensors. The growth of Ag on the hollow nanorings led to rich color variations. Ag-Au nanorings were integrated into ELISA for the sensitive detection of AMD with the naked eye. The proposed method showed high sensitivity for the qualitative and quantitative detection of AMD, the visible cut-off value (0.2 ng mL-1) and limit of detection (0.071 ng mL-1) were 10-fold and 4.7-fold lower, respectively, than those of conventional ELISA. This method showed a linear range of 0.08-2 ng mL-1 and 4-12 ng mL-1. The detection results of this method on 100 samples (food samples and municipal water) agreed well with those of liquid chromatography-tandem mass spectrometry. The proposed plasmonic ELISA has high sensitivity, easy operation, and naked-eye readout.

I2/I--mediated fluorescence quenching of an Ag+-doped gold nanocluster-based immunoassay for sensitive detection of Escherichia coli O157:H7 in milk.

Escherichia coli O157:H7 is a type of hazardous bacteria in the field of food safety. A sensitive and effective method is urgently needed to detect it, avoiding enormous harm for the human health. In this study, we synthesized stable Ag+-doped gold nanoclusters (Ag-AuNC) with a fluorescence intensity 4.8 times stronger than that of AuNC. It was further demonstrated that Ag0 existing in the AuNC core and a fraction of Ag+ anchored on the AuNC shell eliminated the surface defects and improved the luminescent properties of AuNC. A combination of I2 and I- was used to quench fluorescence-enhanced Ag-AuNC, which was first applied in ELISA for detecting E. coli O157:H7 to improve the sensitivity. In the presence of E. coli O157:H7, the biotinylated anti-E. coli O157:H7 mAb and streptavidin-alkaline phosphatase would be immobilized and catalyze l-ascorbic acid 2-phosphate sesquimagnesium salt hydrate to produce ascorbic acid. After addition of KIO3, I2/I- were generated. The I2 could trigger oxidative etching of Ag-AuNC and I- could combine with Ag+ to decrease the Ag+ concentration of Ag-AuNC, which resulted in fluorescence quenching of Ag-AuNC. Under optimal conditions, the linear range of I2/I--mediated fluorescence quenching of Ag-AuNC-based immunoassay for detecting E. coli O157:H7 was 3.3 × 103 to 106 cfu/mL, with a detection limit of 9.2 × 102 cfu/mL, 10.7-fold lower than that of the traditional ELISA. The proposed immunoassay exhibits excellent sensitivity, specificity, recovery, and accuracy, which is useful for quantitative detection of E. coli O157:H7 in food safety.

Development of a label-free plasmonic gold nanoparticles aggregates sensor on the basis of charge neutralization for the detection of zearalenone.

Mycotoxin contamination of corn has been considered a serious problem because it can accumulate in different organs or tissues via ingestion or skin contact and cause several health problems in humans. We have constructed a label-free, colorimetric, and fluorescence dual-channel sensing platform for the detection of zearalenone. Here, we demonstrate that plasmonic gold nanoparticles aggregates could be rapidly formed on the basis of charge neutralization by positively charged SYBR Green I. The sensing platform allowed quantitative detection as low as 0.89 µg kg-1 and visual detection as low as 2.5 µg kg-1. The charge neutralization strategy eliminates a major source of instability in conventional gold nanoparticles colorimetric measurements and paves the way for accurate, label-free bioanalysis.

Dual-mode immunoassay system based on glucose oxidase-triggered Fenton reaction for qualitative and quantitative detection of danofloxacin in milk.

In this study, a novel colorimetric and fluorescent dual-mode ELISA based on glucose oxidase (GOx)-triggered Fenton reaction was developed for the qualitative and quantitative detection of danofloxacin (DAN). In this system, streptavidin-linked biotinylated anti-DAN-monoclonal antibody (SA-Bio-mAb) and biotinylated GOx (Bio-GOx) form the immune complex mAb-Bio-SA-Bio-GOx. In the absence of DAN, the mAb-Bio-SA-Bio-GOx would be immobilized by combining with coated DAN-BSA and catalyzed glucose to generate H2O2. The Fenton reaction between H2O2 and Fe2+ generated hydroxyl radicals, which oxidized the o-phenylenediamine to 2,3-diamino-phenazine. A dual-signal immunoassay with colorimetry and fluorescence as the signal readout was established. In the presence of DAN, DAN and DAN-BSA competed with Bio-mAb, decreasing the connection between immune complexes and DAN-BSA and finally resulting in lower signal of colorimetry and fluorescence. Under optimal conditions, the limit of detection of the fluorescence immunoassay was 0.337 ng/mL and was 5.24-fold lower than that of traditional ELISA. The colorimetric immunoassay cut-off value was 30 ng/mL in milk. The average recoveries of the method for milk samples that are spiked with different concentrations of DAN were 91.1 to 128.3%, with a coefficient of variation of 0.7 to 8.2%. These results of the method exhibited good agreement with those of liquid chromatography-tandem mass spectrometry system (LC-MS/MS) method. In brief, this work provides an improved screening strategy with high sensitivity and accuracy for the qualitative or quantitative detection of DAN in milk monitoring.

Gold nanorods etching-based plasmonic immunoassay for qualitative and quantitative detection of aflatoxin M1 in milk.

In this study, we developed an indirect competitive plasmonic immunoassay using glucose oxidase (GOx)-induced redox reaction to etch Au nanorods (AuNRs) for qualitative and quantitative detection of aflatoxin M1 (AFM1) in milk. In this system, streptavidin (SA) was selected as a linker between biotinylated anti-AFM1-monoantibody (bio-mAb) and biotinylated GOx (bio-GOx) to form the immunocomplexes bio-mAb-SA-bio-GOx. After the oxidation of the glucose and I-, the resultant I2 could etch cetytrimethylammonium bromide (CTAB)-stabilized AuNRs. This resulted in the blue shift of the longitudinal localized surface plasmon resonance (LSPR) extinction peak, with a color change from blue to pink. The linear range and limit of detection (LOD) of the plasmonic immunoassay were 0.25-10 ng mL-1 and 0.11 ng mL-1, respectively. It displayed quantitative detection with high sensitivity, specificity, recovery, and accuracy, which is promising for qualitative and quantitative detection of AFM1 in food safety.

Glucose oxidase-induced colorimetric immunoassay for qualitative detection of danofloxacin based on iron (â ¡) chelation reaction with phenanthroline.

In this study, we developed a competitive colorimetric immunoassay for qualitative detection of DAN based on oxidation of iron (Ⅱ) (Fe2+) in the presence of glucose oxidase (GOx) and color change induced by Fe2+-phenanthroline (Phen) chromogenic system. Streptavidin (SA) acted as a linker between biotinylated anti-DAN-monoantibody (bio-mAb) and biotinylated GOx (bio-GOx) to form the immunocomplexes bio-mAb-SA-bio-GOx. In the absence of DAN, the immunocomplexes bio-mAb-SA-bio-GOx combining with coated DAN-ovalbumin (DAN-OVA) will be immobilized and catalyze glucose to produce H2O2. Fe2+ is oxidized to Fe3+ by H2O2, giving rise to a colorless result. In the presence of DAN, Fe2+ produces a chelation reaction with Phen, leading to orange-red color. Under optimal conditions, the detection limit (LOD) by naked eyes was 2.5 ng mL-1 in milk, chicken, beef, and pork samples. Low LOD, no matrix effect, and no signal reader requirement make it possibly applied to quickly screen DAN on site.

Preparation of an Antidanofloxacin Monoclonal Antibody and Development of Immunoassays for Detecting Danofloxacin in Meat.

Danofloxacin (DAF), a third-generation fluroquinolone (FQ), is widely used as a broad-spectrum antibacterial drug to prevent diseases in livestock and poultry. In this study, a highly specific and sensitive monoclonal antibody (mAb) against DAF was prepared. Also, the mAb was used for the indirect competitive enzyme-linked immunosorbent assay (icELISA) and immunochromatographic strip for the detection of DAF residues in meat. The IC50 of the icELISA based on this mAb was 1.39 ng/mL, and the limit of detection was 0.2 ng/mL. According to the cross-reactivity (CR) experiment, the ELISA that we developed was highly specific and had low CR with other FQ analogues. Moreover, the cut-off of the immunochromatographic strip developed for detecting DAF in meat was 5 ng/mL. Overall, the developed ELISA and immunochromatographic strip based on the prepared mAb were proved reliable for the rapid detection of DAF in meat and can be considered as effective screening methods for food safety and quality management.

Lateral Flow Immunoassay Based on Polydopamine-Coated Gold Nanoparticles for the Sensitive Detection of Zearalenone in Maize.

In this work, polydopamine-coated gold nanoparticles (Au@PDAs) were synthesized by the oxidative self-polymerization of dopamine (DA) on the surface of AuNPs and applied for the first time as a signal-amplification label in lateral flow immunoassays (LFIAs) for the sensitive detection of zearalenone (ZEN) in maize. The PDA layer functioned as a linker between AuNPs and anti-ZEN monoclonal antibody (mAb) to form a probe (Au@PDA-mAb). Compared with AuNPs, Au@PDA had excellent color intensity, colloidal stability, and mAb coupling efficiency. The limit of detection of the Au@PDA-based LFIA (Au@PDA-LFIA) was 7.4 pg/mL, which was 10-fold lower than that of the traditional AuNP-based LFIA (AuNP-LFIA) (76.1 pg/mL). The recoveries of Au@PDA-LFIA were 93.80-111.82%, with the coefficient of variation of 1.08-9.04%. In addition, the reliability of Au@PDA-LFIA was further confirmed by the high-performance liquid chromatography method. Overall, our study showed that PDA coating can chemically modify the surface of AuNPs through a simple method and can thus significantly improve the detection sensitivity of LFIA.

Using molecular descriptors for assisted screening of heterologous competitive antigens to improve the sensitivity of ELISA for detection of enrofloxacin in raw milk.

The use of the heterologous competitive strategy has become a vital method to improve the sensitivity of ELISA. In this work, we prepared an anti-enrofloxacin (ENR) mAb with ENR-bovine serum albumin (BSA) as immunogen. The molecular descriptors of quinolones were then used to screen heterologous coating antigens for the detection of ENR based on an ensemble learning method to improve the sensitivity of the ELISA. Results indicated that 6 of the 7 selected heterologous competitive antigens could enhance the sensitivity of ELISA. The ELISA sensitivity for the detection of ENR with sarafloxacin-BSA as heterologous coating antigen was improved 10-fold (in PBS) and 6-fold (in milk) compared with that with ENR-BSA as homologous antigen. The strategy can effectively screen suitable heterologous competitive antigens to improve the sensitivity of ELISA, followed by preparation of mAb with no additional modification to the corresponding immunogen.