Hendrickson Olga D, Byzova Nadezhda A, Dzantiev Boris B, Zherdev Anatoly V
A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
Foods. 2024 Sep 24;13(19):3032. doi: 10.3390/foods13193032.
and are relevant foodborne bacterial pathogens which may cause serious intoxications and infectious diseases in humans. In this study, a sensitive immunochromatographic analysis (ICA) for the simultaneous detection of these two pathogens was developed. For this, test strips containing two test zones with specific monoclonal antibodies (MAb) against lipopolysaccharides of and and one control zone with secondary antibodies were designed, and the double-assay conditions were optimized to ensure high analytical parameters. Prussian blue nanoparticles (PBNPs) were used as nanozyme labels and were conjugated with specific MAbs to perform a sandwich format of the ICA. Peroxidase-mimic properties of PBNPs allowed for the catalytic amplification of the colorimetric signal on test strips, enhancing the assay sensitivity. The limits of detection (LODs) of and cells were 2 × 10 and 7 × 10 cells/mL, respectively. LODs were 100-fold less than those achieved due to the ICA based on the traditional gold label. The developed double ICA was approbated for the detection of bacteria in cow milk samples, which were processed by simple dilution by buffer before the assay. For and , the recoveries from milk were 86.3 ± 9.8 and 118.2 ± 10.5% and correlated well with those estimated by the enzyme-linked immunosorbent assay as a reference method. The proposed approach was characterized by high specificity: no cross-reactivity with other bacteria strains was observed. The assay satisfies the requirements for rapid tests: a full cycle from sample acquisition to result assessment in less than half an hour. The developed ICA has a high application potential for the multiplex detection of other foodborne pathogens.
[具体细菌名称1]和[具体细菌名称2]是相关的食源性细菌病原体，可导致人类严重中毒和传染病。在本研究中，开发了一种用于同时检测这两种病原体的灵敏免疫层析分析（ICA）方法。为此，设计了包含两个测试区的试纸条，其中测试区含有针对[具体细菌名称1]和[具体细菌名称2]脂多糖的特异性单克隆抗体（MAb），以及一个含有二抗的对照区，并对双检测条件进行了优化，以确保高分析参数。普鲁士蓝纳米颗粒（PBNP）用作纳米酶标记，并与特异性MAb偶联以进行ICA的夹心形式。PBNP的过氧化物酶模拟特性允许催化放大试纸上的比色信号，提高检测灵敏度。[具体细菌名称1]和[具体细菌名称2]细胞的检测限（LOD）分别为2×10和7×10个细胞/mL。与基于传统金标记的ICA相比，LOD降低了100倍。所开发的双ICA方法在检测牛奶样品中的细菌方面得到了验证，检测前牛奶样品通过缓冲液简单稀释进行处理。对于[具体细菌名称1]和[具体细菌名称2]，牛奶中的回收率分别为86.3±9.8%和118.2±10.5%，与作为参考方法的酶联免疫吸附测定法估计的回收率相关性良好。所提出的方法具有高特异性：未观察到与其他细菌菌株的交叉反应。该检测方法满足快速检测的要求：从样品采集到结果评估的整个周期不到半小时。所开发的ICA在其他食源性病原体的多重检测方面具有很高的应用潜力。
