快速检测阪崎肠杆菌新型凝集技术研究

Establishment of a new agglutination technology for rapid detection of Enterobacter sakazakii based on immune colored silica nanoparticles

为建立新型快速检测阪崎肠杆菌(E.sakazakii)的技术,本研究采用有机活性染料与抗体共修饰二氧化硅活性微球制成单分散免疫彩色二氧化硅微球(Immune colored silica nanoparticles,ICSN),建立了快速的E.sakazakii新型凝集检测技术。扫描电镜检测显示,利用反相微乳法制备彩色二氧化硅纳米微球(Colored silica nanoparticles,CSN)大小均匀(约70μm),分散程度好。通过ICSN凝集试验检测E.sakazakii结果表明,该新型凝集技术对目标菌的检测范围为6×103cfu/mL^6×109cfu/mL,其凝集结果清晰、直观、易于判别;重复性和稳定性好,而且具有良好的特异性和准确性。实验结果显示,采用ICSN建立的新型凝集技术具有灵敏、快速、稳定、简便、准确及低成本等优点,适用于快速检测E.sakazakii,并为其它病原性微生物快速检测方法的建立提供技术平台。

To explore a new technology for the rapid detection of Enterobacter sakazakii, we established a new agglutination technique for rapid detection of E.sakazakii by synthesis of the immune colored silica nanoparticles （ICSN） using the organic reactive dyes and coating with the antibody against E.sakazakii. The color silica microspheres （CSN） prepared by microemulsion technique were evenly spheroid with the diameter around 70 μm examined by scanning electron microscopy. The agglutination test results showed that the new agglutination test was specific for E.sakazakii with the detection of 6 ×10^3 cfu/mL and no crossing-reaction with other bacteria. The aggregation results were easy to be judged by naked eyes. In addition, it was stable and repeatable. This new agglutination test could not only be used for rapidly detecting E.sakazakii, but also provide a basis model for rapid detection of other pathogenic bacterium.