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基于双免疫磁珠的侧向流测试条用于同时定量检测癌胚抗原和神经元特异性烯醇化酶。

Dual Immunomagnetic Nanobeads-Based Lateral Flow Test Strip for Simultaneous Quantitative Detection of Carcinoembryonic Antigen and Neuron Specific Enolase.

机构信息

Outpatient Department, Zhujiang Hospital, Southern Medical University, 253 Gongye Road, Guangzhou, Guangdong 510280, China.

School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.

出版信息

Sci Rep. 2017 Feb 10;7:42414. doi: 10.1038/srep42414.

Abstract

A novel immunomagnetic nanobeads -based lateral flow test strip was developed for the simultaneous quantitative detection of neuron specific enolase (NSE) and carcinoembryonic antigen (CEA), which are sensitive and specific in the clinical diagnosis of small cell lung cancer. Using this nanoscale method, high saturation magnetization, carboxyl-modified magnetic nanobeads were successfully synthesized. To obtain the immunomagnetic probes, a covalent bioconjugation of the magnetic nanobeads with the antibody of NSE and CEA was carried out. The detection area contained test line 1 and test line 2 which captured the immune complexes sensitively and formed sandwich complexes. In this assay, cross-reactivity results were negative and both NSE and CEA were detected simultaneously with no obvious influence on each other. The magnetic signal intensity of the nitrocellulose membrane was measured by a magnetic assay reader. For quantitative analysis, the calculated limit of detection was 0.094 ng/mL for NSE and 0.045 ng/mL for CEA. One hundred thirty clinical samples were used to validate the test strip which exhibited high sensitivity and specificity. This dual lateral flow test strip not only provided an easy, rapid, simultaneous quantitative detection strategy for NSE and CEA, but may also be valuable in automated and portable diagnostic applications.

摘要

一种基于免疫磁性纳米珠的新型侧向流测试条被开发出来,用于同时定量检测神经元特异性烯醇化酶(NSE)和癌胚抗原(CEA),这两种标志物在小细胞肺癌的临床诊断中具有高度的敏感性和特异性。利用这种纳米尺度的方法,成功合成了具有高饱和磁化强度、羧基修饰的磁性纳米珠。为了获得免疫磁性探针,通过共价偶联的方式将 NSE 和 CEA 的抗体与磁性纳米珠结合。检测区域包含测试线 1 和测试线 2,它们能够敏感地捕获免疫复合物,并形成三明治复合物。在该测定中,交叉反应结果为阴性,同时检测 NSE 和 CEA,彼此之间没有明显的影响。通过磁测定仪测量硝酸纤维素膜上的磁信号强度。进行定量分析时,计算出 NSE 的检测限为 0.094ng/mL,CEA 的检测限为 0.045ng/mL。该测试条使用 130 份临床样本进行了验证,结果显示其具有较高的灵敏度和特异性。这种双侧向流测试条不仅为 NSE 和 CEA 提供了一种简便、快速、同时定量检测的策略,而且在自动化和便携式诊断应用中可能具有重要价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f4f2/5301198/a37d8d79b635/srep42414-f1.jpg

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