金纳米粒子增敏的SPRi技术应用于遗传性血栓疾病的检测

Au NPs assisted detection of genetic thrombotic diseases by the SPRi technique

目的:基于表面等离子共振成像(SPRi)技术,欲对遗传性血栓疾病4种基因型实现高通量高灵敏的检测。方法:采用多聚腺苷酸(polyA)嵌块将探针简单快速地固定在金表面,制备了polyA-DNA-Au NPs作为增敏介质,结合SPRi技术的检测方法,对遗传性血栓疾病中凝血酶原基因G20210A和凝血因子FV G1691A 4种DNA基因型进行了高通量定量检测。同时,对检测的可行性、灵敏度以及稳定性进行分析研究。结果:4种基因型的检测限可分别低至10.5 pmol/L,12.0 pmol/L,8.0 pmol/L和11.0 pmol/L。在超过16个检测周期后,信号强度仍然保持在98%以上,表明该检测技术的敏感性、重复性较好。结论:金纳米粒子增敏的SPRi技术可实现高通量高灵敏的遗传性血栓疾病标志物检测,并有望推广到更多DNA疾病的检测中。

Aims:Based on the surface plasmon resonance imaging(SPRi)technology,the multiplexing and high sensitivity detection for four genotypes of genetic thrombotic diseases was achieved.Methods:The probe could be fixed to the surface of Au simply and quickly by polyadenylic acid(polyA)blocks;and polyA-DNA-Au NPs were prepared as the medium for enhancing the sensitivity.Combined with the methods of SPRi technology,the multiplexing quantitative detection for four DNA genotypes of prothrombin gene G 20210 A and coagulation factor FV G169A of genetic thrombotic diseases was conducted.Meanwhile,the feasibility,sensitivity,and stability of detection were analyzed and studied.Results:The detection limits of the four genotypes were as low as 10.5 pmol/L,12.0 pmol/L,8.0 pmol/L and 11.0 pmol/L,respectively.After more than 16 detection cycles,the intensity of the signal still remained above 98%,indicating good sensitivity and repeatability of the detection.Conclusions:The SPRi technology sensitized by Au NPs has capability of achieving multiplexing and highly sensitive detection of genetic thrombotic diseases,which is expected to be extended to the detection of more DNA diseases.