Ultra-sensitive method based on time-resolved fluorescence immunoassay for detection of sulfamethazine in raw milk

Song, Huacan; Li, Danhong; Huang, Zhen; Xing, Keyu; Yuan, Chunhong; Peng, Juan; Lai, Weihua

doi:10.1080/09540105.2018.1520816

Cited by 16 publications

(11 citation statements)

References 34 publications

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“…Fifteen real samples for validation experiment were collected from local markets in Nanchang. The detective antigen, SMZ-BSA, was prepared in our lab by coupling SMZ to BSA through diazotization (Hu et al, 2018). All the solvents and other chemicals were of analytical reagent grade.…”

Section: Materials and Reagentsmentioning

confidence: 99%

“…The coupling pH must be optimized because this factor can influence both coupling efficiency and mAb activity. In this section, optimal single key parameters were selected on the basis of two aspects: FI T0 and 1−FI T /FI T0 (Hu et al, 2018). In this study, a high FI T0 and FI inhibition ratio can enhance the sensitivity and broaden the detection range of AIEFM-LFIA.…”

Section: Optimization Of the Coupling Phmentioning

confidence: 99%

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Aggregation-induced emission-based competitive lateral flow immunoassay for rapid detection of sulfamethazine in honey

Wang

Song

Zhang

et al. 2019

Food and Agricultural Immunology

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Sulfamethazine (SMZ), widely used for disease prevention of bees, is harmful to consumers when excess SMZ residues are present in honey. In this study, a novel competitive lateral flow immunoassay (LFIA) was established by using aggregation-induced emission fluorescent microsphere (AIEFM) as label for the detection of SMZ.Results indicated that the limit of detection (LOD) for the developed competitive AIEFM-LFIA in honey was 0.028 ng/mL, and the corresponding linear range was 0.05−5 ng/mL. Recovery experiments showed that the average recovery range was 75.6-95.1% with coefficient variations of 5.7-12.9% in honey. The developed competitive AIEFM-LFIA was compatible with HPLC-MS/MS in detecting SMZ in 15 real samples from honey. In general, the proposed AIEFM-LFIA is well suited for the competitive format and has great potential for the detection of other small targets with single antigenic site in practical application.

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Section: Materials and Reagentsmentioning

confidence: 99%

Section: Optimization Of the Coupling Phmentioning

confidence: 99%

Aggregation-induced emission-based competitive lateral flow immunoassay for rapid detection of sulfamethazine in honey

Wang

Song

Zhang

et al. 2019

Food and Agricultural Immunology

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“…It is necessary to determine the optimal working concentration of Eu 3+ -labelled antibody or antigen in TRFIA (Hu et al, 2018a;Hu et al, 2018b), which not only minimize background interference but also minimize cost investment in the test. As showed in Figure 3, the P/N values of prepared Eu 3+ -labelled goat anti-rabbit IgG monoclonal antibody for anti-Van pAbs in TRFIA were not positively or negatively correlated with its dilution.…”

Section: Preparation Of Eu 3+ -Labelled Goat Anti-rabbit Igg Monoclonmentioning

confidence: 99%

Establishment of an ultrasensitive indirect competitive time-resolved fluoroimmunoassay for vancomycin determination

Han

Dong

et al. 2019

Food and Agricultural Immunology

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Vancomycin (Van) is a common antibiotic that pollutes food, feed and ecological environment. In the present study, an anti-Van polyclonal antibodies (pAbs) and Eu 3+-labeled goat anti-rabbit IgG monoclonal antibody based indirect competitive timeresolved fluoroimmunoassay (IC-TRFIA) was established for ultrasensitive determination of Van, and its half-maximum inhibition concentrations (IC 50) was 13.74 ng/L and the limit of detection (LOD) was 1.07 ng/L. The IC-TRFIA showed strong crossreactivity (CRs) of 36.2% for norvancomycin, and less than 0.1% for polymyxin B, kanamycin and ampicillin. It was found that the IC-TRFIA for Van spiked in different milk powder samples determination were with good accuracy, stability and repeatability, and its recoveries are 83.0-97.9%, also with its coefficient of variations (CVs) are 3.0-12.9% in intra-assay and inter-assay. The results showed that the established IC-TRFIA was promising for ultrasensitive determination of Van in food samples. Highlights (1) Prepared high-quality Van-KLH/BSA conjugates. (2) Obtained high activity and purify anti-Van polyclonal antibodies. (3) Prepared high-quality Eu 3+-labelled goat anti-rabbit IgG monoclonal antibody. (4) Firstly established an IC-TRFIA for Van based on the anti-Van polyclonal antibodies and Eu 3+-labelled goat anti-rabbit IgG monoclonal antibody. (5) The IC-TRFIA can be used for ultrasensitive determination of Van in milk powder samples.

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“…11,12 Although these methods are standardized, accurate and reliable, their wide applications are limited by high costs and complex sample preparations. 13 In addition, antibody-dependent methods including enzyme-linked immunosorbent assay (ELISA), 14 chemiluminescence, 15 time-resolved immunoassay, 16,17 lateral flow immunoassay 18 and electrochemical immunosensor 19 also face different challenges such as poor stabilities, high costs, and difficulties in modifying antibody structures. 20 Aptamers are single-stranded DNA or RNA that bind specifically to target molecules, and they are selected by an in vitro process called systematic evolution of ligands by exponential enrichment (SELEX).…”

Section: Introductionmentioning

confidence: 99%

An Ultrasensitive Label-Free Fluorescent Aptasensor Platform for Detection of Sulfamethazine

Wang¹,

Yan²,

Kou³

et al. 2021

IJN

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Purpose: Sulfamethazine (SMZ) exposed in the environment can enter the human body through the food chain and pose a serious threat to human health. Therefore, it is important to develop a rapid and sensitive method for detecting SMZ in environmental samples. In order to fastly and quantitatively detect SMZ in environmental samples, we developed a label-free fluorescent aptasensor based on specific aptamer (SMZ1S) and fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and rhodamine B (RhoB). Methods: In the absence of SMZ, SMZ1S was adsorbed on the surface of AuNPs, which led to dispersion of the AuNPs in high concentration saline solution, thus effectively quenching the fluorescence of RhoB. With the increase of the SMZ concentration, the specific binding of SMZ1S and SMZ led to the aggregation of AuNPs in the presence of NaCl, which reduced the quenching of RhoB fluorescence and increased the fluorescence intensity. The sensitivity and linearity curve of the label-free fluorescent aptasensor were determined with different concentrations of sulfamethazine standard solutions. The specificity of this fluorescent aptasensor was determined by replacing sulfamethazine with different antibiotics. In addition, the actual water and soil samples were spiked and recovered. Results: Under optimized conditions, the proposed fluorescent aptasensor demonstrated a good linear detection of SMZ in binding buffer from 1.25 ng mL −1 to 40 ng mL −1 and the limit of detection was 0.82 ng mL −1 . The spiked recoveries for SMZ were 94.4% to 108.8% with a relative standard deviation of 1.8-10.3% in water and soil samples, respectively. Conclusion:The label-free fluorescent aptasensor investigated in the current study is a promising tool to detect and quantify SMZ in water and soil samples.

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Ultra-sensitive method based on time-resolved fluorescence immunoassay for detection of sulfamethazine in raw milk

Cited by 16 publications

References 34 publications

Aggregation-induced emission-based competitive lateral flow immunoassay for rapid detection of sulfamethazine in honey

Aggregation-induced emission-based competitive lateral flow immunoassay for rapid detection of sulfamethazine in honey

Establishment of an ultrasensitive indirect competitive time-resolved fluoroimmunoassay for vancomycin determination

An Ultrasensitive Label-Free Fluorescent Aptasensor Platform for Detection of Sulfamethazine

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