Liu Yongcheng, Brandon Robert, Cate Michael, Peng Xiaogang, Stony Robert, Johnson Michael
NN-Labs LLC, P.O. Box 2168, Fayetteville, Arkansas 72702, USA.
Anal Chem. 2007 Nov 15;79(22):8796-802. doi: 10.1021/ac0709605. Epub 2007 Oct 17.
A biosensor system for detection of pathogens was developed by using CdSe/ZnS core/shell dendron nanocrystals with high efficiency and stability as fluorescence labels and a flowing chamber with a microporous immunofilter. The antibody-immobilized immunofilter captured the targeted pathogens, Escherichia coli O157:H7 as an example for bacteria and hepatitis B being a model system for viruses. The CdSe/ZnS core/shell dendron nanocrystals were conjugated with the corresponding antibodies and then passed through the microporous membrane where they attached to the membrane-antigen-antibody. The efficient and stable photoluminescence (PL) of the CdSe/ZnS nanocrystals on the formed "sandwich" structure complexes (membrane-antigen-antibody conjugated with the nanocrystals) was used as the detection means. The effects of the pore size of the membranes, buffer pH, and assay time on the detection of E. coli O157:H7 were investigated and optimized. The detectable level of this new system was as low as 2.3 CFU/mL for E. coli O157:H7 and 5 ng/mL for hepatitis B surface Ag (HBsAg). The assay time was shortened to 30 min without any enrichment and incubation.
通过使用高效稳定的CdSe/ZnS核壳树枝状纳米晶体作为荧光标记物以及带有微孔免疫过滤器的流动腔,开发了一种用于检测病原体的生物传感器系统。固定有抗体的免疫过滤器捕获目标病原体,以大肠杆菌O157:H7作为细菌的示例,以乙型肝炎作为病毒的模型系统。将CdSe/ZnS核壳树枝状纳米晶体与相应抗体偶联,然后使其通过微孔膜,在那里它们附着于膜 - 抗原 - 抗体上。在形成的“三明治”结构复合物(与纳米晶体偶联的膜 - 抗原 - 抗体)上,CdSe/ZnS纳米晶体高效稳定的光致发光(PL)被用作检测手段。研究并优化了膜孔径、缓冲液pH值和检测时间对大肠杆菌O157:H7检测的影响。该新系统对大肠杆菌O157:H7的可检测水平低至每毫升2.3个菌落形成单位(CFU),对乙型肝炎表面抗原(HBsAg)为5纳克/毫升。检测时间缩短至30分钟,无需任何富集和孵育。