Label-free electrochemiluminescence biosensor for ultrasensitive detection of telomerase activity in HeLa cells based on extension reaction and intercalation of Ru(phen)32+
Label-free electrochemiluminescence biosensor for ultrasensitive detection of telomerase activity in HeLa cells based on extension reaction and intercalation of Ru(phen)32+
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基于延伸反应和 Ru(phen)32 嵌入的无标记电化学发光生物传感器超灵敏检测 HeLa 细胞端粒酶活性
DOI：
10.1007/s00216-016-9561-5
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发表时间：
2016-04
影响因子：
4.3
通讯作者：
Guonan Chen
中科院分区：
文献类型：
--
作者：
Bin Qiu;Longhua Guo;Zhenyu Lin;Guonan Chen
端粒酶是人类恶性肿瘤最常见的标志物之一，如子宫、胃、食管、乳腺、结直肠、喉鳞状细胞、甲状腺、膀胱等，因此有必要建立一些灵敏、简便的检测方法来检测端粒酶活性。本研究制备了一种无标记、高灵敏度的电化学发光生物传感器，用于检测HeLa细胞中端粒酶的活性。将巯基化的端粒酶底物（TS）引物通过金-硫（Au-S）相互作用固定在金电极表面，然后用端粒酶特异性地延伸。然后与互补DNA杂交，在电极表面形成双链DNA（dsDNA），Ru（phen）3（2+）嵌入dsDNA的凹槽中，作为电化学发光探针。增强的ECL强度与HeLa细胞数在5 × 5000范围内呈线性关系，检测限为2个HeLa细胞。该电化学发光生物传感器在常见干扰物存在下对端粒酶具有高度的特异性。在100个HeLa细胞上的相对标准偏差（RSD）<5%。该方法为端粒酶相关癌症的筛查或诊断提供了一种方便的方法。
Telomerase is one of the most common markers of human malignant tumors, such as uterine, stomach, esophageal, breast, colorectal, laryngeal squamous cell, thyroid, bladder, and so on. It is necessary to develop some sensitive but convenient detection methods for telomerase activity determination. In this study, a label-free and ultrasensitive electrochemiluminescence (ECL) biosensor has been fabricated to detect the activity of telomerase extracted from HeLa cells. Thiolated telomerase substrate (TS) primer was immobilized on the gold electrode surface through gold-sulfur (Au-S) interaction and then elongated by telomerase specifically. Then, it was hybridized with complementary DNA to form double-stranded DNA (dsDNA) fragments on the electrode surface, and Ru(phen)3 (2+) has been intercalated into the dsDNA grooves to act as the ECL probe. The enhanced ECL intensity has a linear relationship with the number of HeLa cells in the range of 5∼5000 and with a detection limit of 2 HeLa cells. The proposed ECL biosensor has high specificity to telomerase in the presence of common interferents. The relative standard deviations (RSDs) were <5 % at 100 HeLa cells. The proposed method provides a convenient approach for telomerase-related cancer screening or diagnosis.
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影响因子：
14.9
作者：
Huard, S;Moriarty, TJ;Autexier, C
通讯作者：
Autexier, C
影响因子：
56.9
作者：
FENG, JL;FUNK, WD;VILLEPONTEAU, B
通讯作者：
VILLEPONTEAU, B
影响因子：
12.6
作者：
Li, Ying;Liu, Bangwei;Wei, Qingli
通讯作者：
Wei, Qingli
影响因子：
5.4
作者：
Xiaoming Zhou;D. Xing;Debin Zhu;Li Jia
通讯作者：
Xiaoming Zhou;D. Xing;Debin Zhu;Li Jia
影响因子：
13.3
作者：
S. Zong;Zhuyuan Wang;Hui Chen;Yiping Cui
通讯作者：
S. Zong;Zhuyuan Wang;Hui Chen;Yiping Cui