Search

Florfenicol (F), an antimicrobial agent exclusive to veterinary use within the chloramphenicol class, is extensively applied as a broad-spectrum remedy for animal diseases. Despite its efficacy, concerns arise over potential deleterious residues in animal-derived edibles, posing threats to human health. This study pioneers an innovative approach, introducing a quantum dot fluorescence-based immunoassay (FLISA) for the meticulous detection of F residues in animal-derived foods and feeds. This method demonstrates heightened sensitivity, with a detection limit of 0.3 ng/mL and a quantitative detection range of 0.6-30.4 ng/mL. Method validation, applied to diverse food sources, yields recoveries from 90.4 % to 109.7 %, featuring RSDs within 1.3 % to 8.7 %, the results showed high consistency with the national standard HPLC-MS/MS detection method. These findings underscore the method's accuracy and precision, positioning it as a promising tool for swift and reliable F residue detection, with substantial implications for fortifying food safety monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Lomefloxacin (LMF), a third-generation fluoroquinolone antibacterial agent, is often used to treat bacterial and mycoplasma infections. However, due to its prolonged half-life and slow metabolism, it is prone to residues in animal-derived foods, posing a potential food safety risk. Therefore, it is particularly urgent and important to establish a method for detecting lomefloxacin. In this study, direct and indirect competitive fluorescence-linked immunosorbent assay (dc-FLISA and ic-FLISA) based on quantum dots (QDs) was established for the detection of LMF. As for dc-FLISA, the half-maximal inhibitory concentration (IC 50 ) and limit of detection (LOD) were 0.84 ng/mL, 0.04 ng/mL, respectively, the detection ranges from 0.08 to 9.11 ng/mL. The IC 50 and LOD of ic-FLISA were 0.43 ng/mL and 0.03 ng/mL, respectively, meanwhile the detection ranges from 0.05 to 3.49 ng/mL. The recoveries of dc-FLISA and ic-FLISA in animal-derived foods (milk, fish, chicken, and honey), ranged from 95.8% to 105.2% and from 96.3% to 103.4%, respectively, with the coefficients of variation less than 8%. These results suggest that the dc-FLISA and ic-FLISA methods, which are based on QD labelling, are highly sensitive and cost-effective, and can be effectively used to detect LMF in animal-derived foods.

Bacitracin zinc (BAC), a polypeptide antibiotic, is utilized as a feed additive due to its ability to promote growth in animals. However, the abuse of BAC can lead to a great threat to food safety. Therefore, there is an urgent need to develop a rapid and sensitive detection method. In this study, a monoclonal antibody (mAb) against BAC with excellent sensitivity and specificity was obtained. For the first time, quantum dots (QDs) were conjugated with the prepared mAb against BAC and rabbit anti-mouse antibody to fabricate a direct and an indirect competitive fluorescence-linked immunosorbent assay (dc-FLISA and ic-FLISA) to detect BAC. The IC 50 of dc-FLISA and ic-FLISA were 0.28 ng/ml and 0.17 ng/ml, respectively. The limits of detection were 0.0016 ng/ml and 0.001 ng/ml, respectively, and the detection ranges were 0.0016-46.50 ng/ml and 0.001-35.65 ng/ml, respectively. In addition, the recovery rate of the two methods ranged from 93.5% to 112.0%, and the coefficient of variation (CV) was less than 10%. Therefore, the methods developed in this work have the merits of low cost, simple operation, and high sensitivity, which provide an effective analytical tool for BAC residue detection in feed samples., (© 2022 John Wiley & Sons Ltd.)

Based on a highly sensitive and specific monoclonal antibody (mAb) against virginiamycin M1 (VIR M1), a quantum dots-based fluorescent immunochromatographic assay (QDs-ICA) for quick and sensitive analysis of VIR M1 was established for the first time. The mAb showed a half-maximal inhibitory concentration (IC 50 ) of 0.5 ng/mL and cross-reactivity (CR) values below 0.1% for other three analogues when used in enzyme-linked immunosorbent assay (ELISA). The mAb was conjugated to ZnCdSe/ZnS (core/shell) QDs with maximum emission wavelength of 610 nm (orange-red) which was selected as fluorescent probe to increase QDs-ICA sensitivity. The cut-off value of QDs-ICA was 12.5 ng/mL. QDs-ICA showed a linear range from 0.7 to 14.5 ng/mL with a limit of quantification of 0.7 ng/mL. Compared with existing methods for the analysis of VIR M1, the QDs-ICA exhibited higher sensitivity. For analysis of VIR M1 concentrations spiked into swine feed, muscle and liver samples, recovery rates ranged from 94.0% to 111.6% with the highest coefficient of variation (CV) of 6.7% for intra-assay, and for inter-assay ranged from 94.7% to 107.6% with the highest CV of 9.4%. In conclusion, the QDs-ICA could be a potential method for analyzing VIR M1 in animal feed and animal-derived food.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the pathogenic agent leading to COVID-19. Due to high speed of transmission and mutation rates, universal diagnosis and appropriate prevention are still urgently needed. The nucleocapsid protein of SARS-CoV-2 is considered more conserved than spike proteins and is abundant during the virus' life cycle, making it suitable for diagnostic applications. Here, we designed and developed a fluorescent immunochromatography assay (FICA) for the rapid detection of SARS-CoV-2-specific antibodies using ZnCdSe/ZnS QDs-conjugated nucleocapsid (N) proteins as probes. The nucleocapsid protein was expressed in E.coli and purified via Ni-NTA affinity chromatography with considerable concentration (0.762 mg/mL) and a purity of more than 90%, which could bind to specific antibodies and the complex could be captured by Staphylococcal protein A (SPA) with fluorescence displayed. After the optimization of coupling and detecting conditions, the limit of detection was determined to be 1:1.024 × 10 5 with an IgG concentration of 48.84 ng/mL with good specificity shown to antibodies against other zoonotic coronaviruses and respiratory infection-related viruses ( n = 5). The universal fluorescent immunochromatography assay simplified operation processes in one step, which could be used for the point of care detection of SARS-CoV-2-specific antibodies. Moreover, it was also considered as an efficient tool for the serological screening of potential susceptible animals and for monitoring the expansion of virus host ranges.

African swine fever (ASF), a highly contagious and lethal disease, poses a tremendous threat and burden to the swine industry worldwide. Lack of available vaccines or treatments leaves rapid diagnosis as the key tool to control the disease. Quantum dots (QDs) are unique fluorescent semiconductor nanoparticles, highly versatile for biological applications. In this study, we developed a quantum dots-based fluorescent immunochromatographic assay (QDs-FICA) using CD2v as the diagnosis antigen to detect ASFV antibodies. The titre of the test strip was 1 : 5·12 × 10 5 . In addition, the strip was highly specific to anti-ASFV serum and had no cross-reaction with CSFV, PPV, PRRSV, PCV-2, PRV and FMDV. Moreover, a comparative test of 71 clinical samples showed that the coincidence rate was 85·92% between the test strip and the commercial ELISA kit (coated with p30, p62 and p72). The QDs-FICA can be used to detect ASFV antibodies, which is meaningful for the surveillance, control and purification of ASF., (© 2022 The Society for Applied Microbiology.)

Arsanilic acid (ASA) residue, which is the most common contaminant in edible animal tissues such as pork and liver, has caused environmental and food-safety concerns. In this study, direct and indirect competitive fluorescence-linked immunosorbent assays (dc-FLISA and ic-FLISA) incorporating quantum dots (QDs) as the fluorescent label were developed for the first time to detect ASA residues in edible pork and animal liver. Monoclonal antibodies against ASA and rabbit anti-mouse antibody were conjugated to orange QDs with excitation wavelengths at 450 nm, and the QD-Abs served as detection probes. The limits of detection for dc-FLISA and ic-FLISA were 0.11 ng/mL and 0.001 ng/mL, respectively. QD-FLISA was used to analyse spiked samples; recoveries ranged from 80.2%-91.2% in dc-FLISA and 82.5%-91.2% in ic-FLISA, and the coefficients of variations (CV) were less than 12%. Compared with conventional indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), the QD-FLISA described here was more sensitive and accurate in the analysis of ASA residues in animal tissues. Moreover, the results of QD-FLISA correlated well with HPLC. These results indicate that dc-FLISA and ic-FLISA are sensitive and reliable for detection of ASA residues in edible animal tissues.

Ochratoxin A (OTA) is a toxic metabolite produced mainly by Aspergillus and Penicillium species. A quantitative method was developed for the rapid, simple, and sensitive detection of OTA in corn by quantum dots-based fluorescent immunochromatographic assay (QDs-ICA). The CdSe/ZnS QDs-labelled anti-OTA monoclonal antibody (mAb) conjugates were designed as the fluorescent signal probe. The QDs-ICA included the designation of test line (T line) and control line (C line), which were sprayed with optimal concentrations of the OTA-OVA and staphylococcal protein-A (SPA), respectively. Under the optimal experimental conditions, the QDs-ICA exhibited excellent specificity and good accuracy and precision. For qualitative detection, the cut-off value for the T line of the visual detection method was 2.5 ng/mL. For quantitative detection, the linear regression equation of the standard curve was y = 0.366x + 0.514 with a reliable correlation coefficient (R 2  = 0.992). Moreover, the 50% inhibition value (IC 50 ) of the QDs-ICA was 0.91 ng/mL, the limit of detection (LOD) was 0.07 ng/mL, and the detection range was 0.05 to 10 ng/mL. In addition, the recovery rates ranged from 91.82 to 100.35% with a coefficient of variation (CV) below 3% for intra-assay, whereas the recovery rates for inter-assay changed from 94.29 to 104.62% with a CV below 10%. These results indicate that the QDs-ICA can serve as a potential large-scale preliminary device for rapid determination of OTA. Using CdSe/ZnS QDs as the fluorescent signal for quantum dots-based fluorescent immunochromatographic assay, the QDs-ICA provided a novel method for the rapid simultaneous qualitative and quantitative determination of OTA.

The direct and indirect competitive fluorescence-linked immunosorbent assay (FLISA and icFLISA) incorporating quantum dots (QDs) for the detection of fleroxacin (FLE) was established for the first time in this study. The monoclonal antibody specific for FLE was successfully conjugated with QDs after purification by the caprylic acid-ammonium sulphate method. The limits of detection of FLISA and icFLISA were 0.012 ng/mL and 0.006 ng/mL, respectively; IC50 were 0.32 ng/mL and 0.19 ng/mL; and the detection ranges were 0.012–24.490 ng/mL and 0.006–16.210 ng/mL. The recovery was 93.8%-112.4% and the coefficient of variation was below 11.75%. The fabricated FLISA and icFLISA are cost-effective, high sensitive and can be an alternative method in the detection of FLE residues. [ABSTRACT FROM AUTHOR]

A sensitive and accurate multiple fluorescence immunoassay for the simultaneous quantitative detection of Zearalenone (ZEN) and Ochratoxin A (OTA) in single spot based on multicolor quantum dots (QDs) labeling was developed for the first time. Two kinds of ZnCdSe/ZnS (core/shell) QDs with maximum emission wavelengths at 520 nm (green) and 610 nm (orange-red) were selected as marking materials, respectively. The anti-ZEN-mAb-QDs and anti-OTA-mAb-QDs were designed as the immune fluorescent probes. Fluorescence was measured at the same excitation wavelength and two different emission wavelengths to determine each target. The procedure for QDs-based multiple fluorescence labeled immunosorbent assay (M-FLISA) was developed. The 50% inhibition concentrations (IC 50) of ZEN and OTA were 0.034 and 1.175 ng/mL. Moreover, the limits of detection (LOD) for the simultaneous determination were 0.0239 and 2.339 ng/g for ZEN and OTA in maize, respectively. In addition, the recoveries ranged from 93.15 to 101.90% for ZEN and from 95.29 to 102.43% for OTA, with the coefficient variation (CV) of 2.70–8.86% and 3.51–6.22% respectively. There was good consistency between the M-FLISA and high performance liquid chromatography (HPLC) results, which confirmed that the M-FLISA was suitable for the simultaneous quantitative detection of various mycotoxins. The schematic diagram of simultaneous determination of ZEN and OTA based on multicolor quantum dots. [Display omitted] • Two kinds of quantum dots with different emission wavelengths were selected as labels. • Simultaneous detection of Zearalenone and Ochratoxin A based on quantum dots labeled antibody. • This multiple fluorescence immunoassay for the detection possessed high sensitivity and perfect accuracy. • Simultaneous quantitative detection of Zearalenone and Ochratoxin A in a single well were developed for the first time. [ABSTRACT FROM AUTHOR]