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基于表面增强拉曼散射的侧向流动免疫分析中使用纳米银聚多巴胺纳米球同时检测宫颈癌血清中的鳞状细胞癌抗原和癌抗原125 。

The simultaneous detection of the squamous cell carcinoma antigen and cancer antigen 125 in the cervical cancer serum using nano-Ag polydopamine nanospheres in an SERS-based lateral flow immunoassay.

作者信息

Xia Ji, Liu Yifan, Ran Menglin, Lu Wenbo, Bi Liyan, Wang Qian, Lu Dan, Cao Xiaowei

机构信息

Department of Obstetrics and Gynecology, College of Clinical Medicine, Yangzhou University Yangzhou PR China

The First Clinical College, Dalian Medical University Dalian PR China.

出版信息

RSC Adv. 2020 Aug 6;10(49):29156-29170. doi: 10.1039/d0ra05207h. eCollection 2020 Aug 5.

Abstract

The accurate analysis of tumor related biomarkers is extremely critical in the diagnosis of the early stage cervical cancer. Herein, we designed a novel and inexpensive surface-enhanced Raman scattering-based lateral flow assay (SERS-based LFA) strip with a single test line, which was applied for the rapid and sensitive quantitative simultaneous analysis of SCCA and CA125 in serum samples from patients with cervical cancer. In the presence of target antigens, the monoclonal antibody-coupled and Raman reporter-labeled nano-Ag polydopamine nanospheres (PDA@Ag-NPs) aggregated on the test line modified by the polyclonal antibody to form a double-antibody sandwich structure. The finite difference time domain simulation demonstrated that large number of "hot spots" was generated among the nanogaps of aggregated PDA@AgNPs, which resulted in a huge enhancement of the signal of the Raman reporters. Accordingly, the limit of detection was determined to be 7.156 pg mL for SCCA and 7.182 pg mL for CA125 in phosphate buffer and 8.093 pg mL for SCCA and 7.370 pg mL for CA125 in human serum, revealing high sensitivity of this SERS-based LFA strip. Significantly, the detection of SCCA and CA125 using the SERS-based LFA was observed to have high specificity and reproducibility, and the whole detection was completed within 20 min. Furthermore, the SERS-based LFA and enzyme-linked immunosorbent assay were also employed in serum samples obtained from patients with cervical cancer, cervical intraepithelial neoplasia and healthy subjects, and perfect agreement existed between both the methods. Thus, clinically, the developed SERS-based LFA strip has strong potential for the simultaneous detection of multiple cancer biomarkers in serum.

摘要

肿瘤相关生物标志物的准确分析在早期宫颈癌的诊断中极其关键。在此,我们设计了一种新型且廉价的基于表面增强拉曼散射的侧向流动检测(SERS-LFA)试纸条,其带有一条检测线,用于对宫颈癌患者血清样本中的鳞状细胞癌抗原(SCCA)和癌抗原125(CA125)进行快速、灵敏的定量同步分析。在存在靶抗原的情况下,与单克隆抗体偶联且用拉曼报告分子标记的纳米银聚多巴胺纳米球(PDA@Ag-NPs)在由多克隆抗体修饰的检测线上聚集,形成双抗体夹心结构。时域有限差分模拟表明,聚集的PDA@AgNPs的纳米间隙中产生了大量“热点”,这导致拉曼报告分子的信号大幅增强。因此,在磷酸盐缓冲液中,SCCA的检测限为7.156 pg/mL,CA125的检测限为7.182 pg/mL;在人血清中,SCCA的检测限为8.093 pg/mL,CA125的检测限为7.370 pg/mL,这表明该基于SERS的LFA试纸条具有高灵敏度。值得注意的是,使用基于SERS的LFA检测SCCA和CA125具有高特异性和可重复性,整个检测在20分钟内完成。此外,基于SERS的LFA和酶联免疫吸附测定也用于从宫颈癌、宫颈上皮内瘤变患者和健康受试者获得的血清样本,两种方法之间存在完美的一致性。因此,在临床上,所开发的基于SERS的LFA试纸条在同时检测血清中的多种癌症生物标志物方面具有强大的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1daa/9055935/5fbd47074f46/d0ra05207h-s1.jpg

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