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基于磁致荧光淬灭性能的双模态免疫层析检测技术初探
The Primary Exploration of A Dual-mode Immune Chromatography Testing Technology Based on Magnetic Fluorescence Quenching Performance
【作者】 张博；
【作者基本信息】 天津大学 ， 生物学， 2018， 硕士
【摘要】 免疫层析试纸条检测技术具有操作简便、价格低廉、检测快速、可现场即时检测的优势,是目前传统检测法和仪器检测法的良好补充。然而,传统免疫层析试纸条检测技术由于检测灵敏度低,难以定量等缺点限制了其在医学检测中的发展。针对上述难题,本文将传统胶体金试纸条进行快速可视化定性的特性与荧光检测高灵敏定量的优势相结合,构建出同时具有显色和荧光双信号读出机制的免疫层析试纸条检测技术,具体工作如下:利用超声乳化法成功制备多包裹磁纳米球,将其作为信号标记物,构建了基于磁致荧光淬灭的双模态联检免疫层析试纸条,并全面评价了试纸条的质量与性能。通过磁纳米颗粒的高效荧光淬灭特性和超顺磁性富集和分离待检样品,实现对血清及全血中生物标志物的高灵敏双模态联合检测。结果表明:多包裹磁球荧光淬灭能力强,稳定性好,在对乳腺癌相关标志物CA153和CEA的联合检测中,灵敏度分别可达到0.06 ng/mL和0.09 U/mL,在对急性心肌梗死(AMI)相关标志物cTnI和CK-MB的联合检测中,灵敏度分别可达到0.049 ng/mL和0.089ng/mL,且在实际样本检测中,检测结果与临床常规检测方法一致性良好。综上所述,本文成功构建了基于磁致荧光淬灭的双模态联检免疫层析检测技术,该联检技术提高了肿瘤和急性病症的检出率,降低了成本和样本用量,缩短了检测时间。以此试纸条为平台,今后可推广应用到多种疾病标志物的检测。
【Abstract】 Immunochromatography test strip(ICTS) technology has the advantage of simple,low cost,rapid,and can be realized instant point of care testing(POCT).It is a good supplement to traditional detection and instrument detection method.However,low sensitivity and nonquantitative of traditional ICTS limit its development in clinic testing.In response to the above problems,in this work,we described a magnetic nanosphere fluorescence quenching ICTS(mFICTS)with fluorescence and colorimetric dual signal readout mechanism.The details are as follows:The magnetic nanosphere was successfully prepared by ultrasonic emulsification.Here,we used the magnetic nanospheres probe as signal markers,and prepared the mFICTS.The quality and performance of the mFICTS were comprehensive evaluated.Meanwhile,the samples were enriched and separated by the superparamagnetic of magnetic nanosphere,a high sensitive dual-mode combined detection of biomarkers in serum or even in whole blood was achieved.The results showed that the magnetic nanospheres had strong fluorescence quenching ability and good stability.In the combined detection of breast cancer related markers CA153 and CEA,the detection of limits(LOD)are 0.06 ng/mL and 0.09 U/mL,and for the AMI related markers cTnI and CK-MB,the LOD are 0.049 ng/mL and 0.089 ng/mL,the result is consistent with the clinical detection methods.In conclusion,a dual mode,high sensitive and visual combined ICTS detection technology based on magnetic nanospheres fluorescence quenching is successfully constructed.This combined detection technology improves the detection rate of tumors and other acute diseases,significantly reduces the cost and sample volume,and shortens the detection time,it can be applied to the detection of multiple markers in the future.