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基于适配体偶联纳米粒子的免疫层析条生物传感器快速检测 N-羟乙酰神经氨酸

Immunochromatographic strip biosensor for the rapid detection of N-glycolylneuraminic acid based on aptamer-conjugated nanoparticle.

机构信息

Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, 130062, PR China.

Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, 130062, PR China.

出版信息

Anal Biochem. 2018 Nov 15;561-562:52-58. doi: 10.1016/j.ab.2018.07.017. Epub 2018 Jul 20.

DOI:10.1016/j.ab.2018.07.017
PMID:30036498
Abstract

N-glycolylneuraminic acid (Neu5Gc) is a type of sialic acid that is not typically produced in healthy humans but detective in some visceral cancer cells. As a new carcinoma biomarker, the level change in the serum and urine from the patient could potentially have the relation to the disease progression. So the measurement of the Neu5Gc will help to take a better response to therapeutic schedule for the sufferers. A sensitive and rapid aptamer-nanoparticle immunochromatographic strip for the visual detection of Neu5Gc was developed. The assay is based on the competitive reaction of binding the DNA aptamer targeting the candidate molecule selected by SELEX between Neu5Gc and complementary DNA. The sensing results indicated that the aptamer-based strip was sufficiently sensitive to detect Neu5Gc. The visual limit of detection (LOD) for semi-quantitative detection was 30 ng/mL under the optimal conditions and a quantitative detection limit of 5.38 ng/mL could be obtained using a scanning strip reader. The average recovery of the spiked cancer cell samples was 88.86%, with a coefficient of variation (CV) of 5.27%. The detection could be performed in less than 15 min using a simple procedure without any complicated equipment, demonstrating that this aptamer-nanoparticle biosensor strip has great potential for use to Neu5Gc-related cancer diagnosis.

摘要

N-羟乙酰神经氨酸(Neu5Gc)是一种唾液酸,在健康人体中通常不会产生,但在一些内脏癌细胞中存在。作为一种新的癌标志物,患者血清和尿液中水平的变化可能与疾病的进展有关。因此,Neu5Gc 的测量将有助于更好地应对患者的治疗方案。开发了一种用于 Neu5Gc 可视化检测的灵敏、快速适体-纳米颗粒免疫层析条。该测定法基于通过 SELEX 选择的候选分子与互补 DNA 之间结合针对该候选分子的 DNA 适体的竞争反应。检测结果表明,基于适体的条带对 Neu5Gc 具有足够的灵敏度。在最佳条件下,半定量检测的视觉检测限(LOD)为 30ng/mL,使用扫描条带读取器可获得 5.38ng/mL 的定量检测限。用该方法对添加的癌细胞样本进行检测的平均回收率为 88.86%,变异系数(CV)为 5.27%。该检测可在不到 15 分钟的时间内完成,操作简单,无需任何复杂的设备,表明该适体-纳米颗粒生物传感器条带在 Neu5Gc 相关癌症诊断中具有很大的应用潜力。

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