Key Laboratory of Genetics and Biotechnology, College of Life Sciences, Capital Normal University, Beijing, People's Republic of China.
Appl Microbiol Biotechnol. 2010 Apr;86(3):983-90. doi: 10.1007/s00253-009-2417-6. Epub 2010 Jan 21.
Traditional methods for identifying food-borne pathogens are time-consuming and laborious, so it is necessary to develop innovative methods for the rapid identification of food-borne pathogens. Here, we report the development of silicon-based optical thin-film biosensor chips for sensitive detection of 11 food-borne pathogens. Briefly, aldehyde-labeled probes were arrayed and covalently attached to a hydrazine-derivatized chip surface, and then, biotinylated polymerase chain reaction (PCR) amplicons were hybridized with the probes. After washing and brief incubation with an antibiotin immunoglobulin G-horseradish peroxidase conjugate and a precipitable horseradish peroxidase substrate, biotinylated chains bound to the probes were visualized as a color change on the chip surface (gold to blue/purple). Highly sensitive and accurate examination of PCR fragment targets can be completed within 30 min. This assay is extremely robust, sensitive, specific, and economical and can be adapted to different throughputs. Thus, a rapid, sensitive, and reliable technique for detecting 11 food-borne pathogens was successfully developed.
传统的食源性病原体鉴定方法既费时又费力,因此有必要开发创新的方法来快速鉴定食源性病原体。在这里,我们报告了硅基光学薄膜生物传感器芯片的开发,用于灵敏检测 11 种食源性病原体。简而言之,醛基标记的探针被排列并共价连接到肼衍生的芯片表面,然后,生物素化聚合酶链反应 (PCR) 扩增子与探针杂交。洗涤后,用抗生物素 IgG-辣根过氧化物酶缀合物和可沉淀的辣根过氧化物酶底物短暂孵育后,结合到探针上的生物素化链在芯片表面上显示为颜色变化(金变蓝/紫)。可以在 30 分钟内完成对 PCR 片段靶标的高度敏感和准确检查。该检测方法非常稳健、灵敏、特异且经济实惠,可适用于不同的通量。因此,成功开发了一种快速、灵敏和可靠的检测 11 种食源性病原体的技术。