研究目的
Developing an anti-ovalbumin antibody-based lateral flow immunoassay (LFI) for T-2 toxin detection with improved sensitivity and universality.
研究成果
The anti-OVA antibody-based QB-LFI system successfully combined sensitivity and universality for T-2 toxin detection, with a LOD of 10 fg mL?1. The system's performance was validated through comparison with traditional LFI systems and a commercial ELISA kit, demonstrating its potential for rapid, sensitive, quantitative, and universal detection in biosafety monitoring.
研究不足
The introduced preincubation increased the complexity of the method, requiring strict timing for every step. The method's specificity was tested against structural analogs and other toxins, but potential cross-reactivity with other substances not tested could be a limitation.
1:Experimental Design and Method Selection:
The study employed a lateral flow immunoassay (LFI) using CdSe/ZnS quantum dot beads (QBs) as labels for the detection of T-2 toxin. The method introduced preincubation to improve detection limits and used a coating antigen as a bifunctional element for universality.
2:Sample Selection and Data Sources:
T-2 toxin standard and its metabolites, along with other common toxins, were used to evaluate the specificity of the QB-LFI system. Tap water samples spiked with T-2 toxin were used to test the method's applicability.
3:List of Experimental Equipment and Materials:
Carboxylate-modified QBs, nitrocellulose membrane, glass fiber, absorbent pad, polyvinylchloride backing card, and various reagents were used. Instruments included a Milli-Q purification system, BioJet Quanti 3000?, and a QB-LFI strip reader.
4:Experimental Procedures and Operational Workflow:
The QB surface was modified with anti-T-2 monoclonal antibodies. The QB-LFI system was fabricated with anti-OVA antibody and goat anti-mouse IgG antibodies immobilized on the NC membrane. The quantitative procedure involved preincubation of T-2 toxin and T-2-OVA with QB-mAb, followed by detection on the LFI strip.
5:Data Analysis Methods:
Fluorescence intensity was measured at 610 nm. A calibration plot was established, and the limit of detection (LOD) was determined.
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