Zhang Pengfei, Bao Yan, Draz Mohamed Shehata, Lu Huiqi, Liu Chang, Han Huanxing
Center for Translational Medicine, Changzheng Hospital, Second Military Medical University, Shanghai, People's Republic of China.
Zhejiang-California International Nanosystems Institute, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China ; Faculty of Science, Tanta University, Tanta, Egypt.
Int J Nanomedicine. 2015 Sep 30;10:6161-73. doi: 10.2147/IJN.S89307. eCollection 2015.
Convenient and rapid immunofiltration assays (IFAs) enable on-site "yes" or "no" determination of disease markers. However, traditional IFAs are commonly qualitative or semi-quantitative and are very limited for the efficient testing of samples in field diagnostics. Here, we overcome these limitations by developing a quantum dots (QDs)-based fluorescent IFA for the quantitative detection of C-reactive proteins (CRP). CRP, the well-known diagnostic marker for acute viral and bacterial infections, was used as a model analyte to demonstrate performance and sensitivity of our developed QDs-based IFA. QDs capped with both polyethylene glycol (PEG) and glutathione were used as fluorescent labels for our IFAs. The presence of the surface PEG layer, which reduced the non-specific protein interactions, in conjunction with the inherent optical properties of QDs, resulted in lower background signal, increased sensitivity, and ability to detect CRP down to 0.79 mg/L with only 5 µL serum sample. In addition, the developed assay is simple, fast and can quantitatively detect CRP with a detection limit up to 200 mg/L. Clinical test results of our QD-based IFA are well correlated with the traditional latex enhance immune-agglutination aggregation. The proposed QD-based fluorescent IFA is very promising, and potentially will be adopted for multiplexed immunoassay and in field point-of-care test.
便捷快速的免疫过滤分析(IFA)能够在现场对疾病标志物进行“是”或“否”的判定。然而,传统的IFA通常是定性或半定量的,在现场诊断中对样品进行高效检测的能力非常有限。在此,我们通过开发一种基于量子点(QD)的荧光IFA来定量检测C反应蛋白(CRP),从而克服了这些局限性。CRP是急性病毒和细菌感染的著名诊断标志物,被用作模型分析物来证明我们开发的基于QD的IFA的性能和灵敏度。用聚乙二醇(PEG)和谷胱甘肽包覆的量子点被用作我们IFA的荧光标记物。表面PEG层的存在减少了非特异性蛋白质相互作用,结合量子点固有的光学特性,导致背景信号更低、灵敏度提高,并且仅用5微升血清样品就能检测低至0.79毫克/升的CRP。此外,所开发的检测方法简单、快速,能够定量检测CRP,检测限高达200毫克/升。我们基于量子点的IFA的临床测试结果与传统乳胶增强免疫凝集反应具有良好的相关性。所提出的基于量子点的荧光IFA非常有前景,并且有可能被用于多重免疫分析和现场即时检测。