Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk
<i>Salmonella typhimurium</i> and <i>Listeria monocytogenes</i> 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 o...
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MDPI AG
2024-09-01
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| Series: | Foods |
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| author | Olga D. Hendrickson Nadezhda A. Byzova Boris B. Dzantiev Anatoly V. Zherdev |
| author_facet | Olga D. Hendrickson Nadezhda A. Byzova Boris B. Dzantiev Anatoly V. Zherdev |
| author_sort | Olga D. Hendrickson |
| collection | DOAJ |
| description | <i>Salmonella typhimurium</i> and <i>Listeria monocytogenes</i> 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 <i>S. typhimurium</i> and <i>L. monocytogenes</i> 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 <i>Salmonella</i> and <i>Listeria</i> cells were 2 × 10<sup>2</sup> and 7 × 10<sup>3</sup> 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 <i>S. typhimurium</i> and <i>L. monocytogenes</i>, 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. |
| format | Article |
| id | doaj-art-ff923f05475d4cbf845ef16c7f848e6e |
| institution | OA Journals |
| issn | 2304-8158 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | MDPI AG |
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| series | Foods |
| spelling | doaj-art-ff923f05475d4cbf845ef16c7f848e6e2025-08-20T02:16:50ZengMDPI AGFoods2304-81582024-09-011319303210.3390/foods13193032Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in MilkOlga D. Hendrickson0Nadezhda A. Byzova1Boris B. Dzantiev2Anatoly V. Zherdev3A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia<i>Salmonella typhimurium</i> and <i>Listeria monocytogenes</i> 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 <i>S. typhimurium</i> and <i>L. monocytogenes</i> 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 <i>Salmonella</i> and <i>Listeria</i> cells were 2 × 10<sup>2</sup> and 7 × 10<sup>3</sup> 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 <i>S. typhimurium</i> and <i>L. monocytogenes</i>, 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.https://www.mdpi.com/2304-8158/13/19/3032<i>Salmonella typhimurium</i><i>Listeria monocytogenes</i>foodborne pathogensmultiplex immunochromatographic analysisPrussian blue nanozymefood safety |
| spellingShingle | Olga D. Hendrickson Nadezhda A. Byzova Boris B. Dzantiev Anatoly V. Zherdev Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk Foods <i>Salmonella typhimurium</i> <i>Listeria monocytogenes</i> foodborne pathogens multiplex immunochromatographic analysis Prussian blue nanozyme food safety |
| title | Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk |
| title_full | Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk |
| title_fullStr | Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk |
| title_full_unstemmed | Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk |
| title_short | Prussian-Blue-Nanozyme-Enhanced Simultaneous Immunochromatographic Control of Two Relevant Bacterial Pathogens in Milk |
| title_sort | prussian blue nanozyme enhanced simultaneous immunochromatographic control of two relevant bacterial pathogens in milk |
| topic | <i>Salmonella typhimurium</i> <i>Listeria monocytogenes</i> foodborne pathogens multiplex immunochromatographic analysis Prussian blue nanozyme food safety |
| url | https://www.mdpi.com/2304-8158/13/19/3032 |
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