Aptamer-based microchip electrophoresis assays for amplification detection of carcinoembryonic antigen.
Aptamer-based microchip electrophoresis assays for amplification detection of carcinoembryonic antigen.
复制标题
基于适体的微芯片电泳检测癌胚抗原的扩增检测
DOI：
10.1016/j.cca.2015.09.002
复制
发表时间：
2015-10-23
期刊：
影响因子：
--
通讯作者：
Liu YM
中科院分区：
文献类型：
--
作者：
Pan L;Zhao J;Huang Y;Zhao S;Liu YM
癌胚抗原（CEA）作为最广泛使用的肿瘤标志物之一，用于结直肠癌、胰腺癌、胃癌和宫颈癌的临床诊断。我们开发了一种基于适体的微芯片电泳检测技术，用于检测人血清中的 CEA 以进行癌症诊断。磁珠（MB）用作双链DNA的载体，双链DNA由目标适体和适体的互补DNA形成。 MB-dsDNA 缀合物中的适体与靶标结合后，互补 DNA 从 MB-dsDNA 缀合物中释放出来。释放的互补DNA与荧光素酰胺(FAM)标记的DNA杂交，形成DNA双链体，通过内切核酸酶Nb.BbvCI的切口触发FAM标记的DNA的选择性切割，并生成FAM标记的DNA片段。释放的互补DNA与另一个FAM标记的DNA杂交，导致FAM标记的DNA不断裂解，并产生大量FAM标记的DNA片段。在 MCE 激光诱导荧光检测 (LIF) 中，FAM 标记的 DNA 片段被分离和检测。 CEA的线性范围为130 pg/ml~8.0 ng/ml，相关系数为0.9916，检出限为68 pg/ml。健康受试者血清样本中的 CEA 浓度在 1.3 ng/ml 至 3.2 ng/ml 范围内。癌症患者样本中的 CEA 浓度>15 ng/ml。该方法可能成为生物医学分析和临床诊断中快速分析CEA和其他肿瘤标志物的有用工具。
Carcinoembryonic antigen (CEA) as one of the most widely used tumor marker is used in the clinical diagnosis of colorectal, pancreatic, gastric, and cervical carcinomas. We developed an aptamer-based microchip electrophoresis assay technique for assaying CEA in human serum for cancer diagnosis. The magnetic beads (MBs) are employed as carriers of double strand DNA that is formed by an aptamer of target and a complementary DNA of aptamer. After the aptamer in MB-dsDNA conjugate binds with target, the complementary DNA was released from MB-dsDNA conjugate. The released complementary DNA hybridizes with a fluorescein amidite (FAM) labeled DNA, and forms DNA duplex, which triggers the selective cleavage of FAM labeled DNA by nicking endonuclease Nb.BbvCI, and generating FAM labeled DNA segment. The released complementary DNA hybridizes with another FAM labeled DNA, resulting in a continuous cleavage of FAM labeled DNA, and the generation of large numbers of FAM labeled DNA segments. In MCE laser induced fluorescence detection (LIF), FAM labeled DNA segment is separated and detected. The linear range for CEA was 130 pg/ml~8.0 ng/ml with a correlation coefficient of 0.9916 and a detection limit of 68 pg/ml. The CEA concentration in the serum samples from healthy subjects was found be in the range 1.3 ng/ml to 3.2 ng/ml. The CEA concentration in the samples from cancer patients was found to be >15 ng/ml. This method may become a useful tool for rapid analysis of CEA and other tumor markers in biomedical analysis and clinical diagnosis.
影响因子：
2.9
作者：
Fredlake, Christopher P.;Hert, Daniel G.;Root, Brian E.;Barron, Annelise E.
通讯作者：
Barron, Annelise E.