Fluorescent Biosensor Based on Hairpin DNA Stabilized Copper Nanoclusters for Chlamydia trachomatis Detection
Fluorescent Biosensor Based on Hairpin DNA Stabilized Copper Nanoclusters for Chlamydia trachomatis Detection
复制标题
基于发夹 DNA 稳定铜纳米簇的荧光生物传感器用于沙眼衣原体检测
DOI：
10.1007/s10895-022-02961-y
复制
发表时间：
2022-05-25
影响因子：
2.7
通讯作者：
Chen，Lili
中科院分区：
文献类型：
--
作者：
Liu，Luyao;Bai，Qinqin;Chen，Lili
沙眼衣原体是一种细胞内寄生微生物，可引起沙眼等多种疾病。在该策略中，设计了一种以探针环为特异区域识别沙眼衣原体DNA的特异性发夹DNA，其茎由24个AT碱基对组成，作为发夹DNA- cuncs形成的有效模板。在沙眼衣原体DNA缺失的情况下，检测系统在606 nm处显示出较强的橙色荧光发射峰。在沙眼衣原体DNA存在下，与沙眼衣原体DNA杂交后，DNA探针的构象发生了变化。然后，发夹DNA-CuNCs的数量减少，导致低荧光发射。沙眼衣原体DNA与特定的发夹DNA竞争，对荧光发夹DNA- cuncs的合成有明显的抑制作用。在最佳实验条件下，对不同浓度沙眼衣原体进行检测，结果在50 nM ~ 950 nM范围内呈良好的线性关系。检测限为18.5 nM，具有良好的选择性。最后，该荧光生物传感器成功应用于HeLa细胞裂解液中沙眼衣原体靶序列的检测，为沙眼衣原体的检测提供了新的策略。
Chlamydia trachomatis (C. trachomatis) is a kind of intracellular parasitic microorganism, which can causes many diseases such as trachoma. In this strategy, a specific hairpin DNA with the probe loop as specific regions to recognize C. trachomatis DNA with strong affinity was designed, and its stem consisted of 24 AT base pairs as an effective template for hairpin DNA-CuNCs formation. In the absence of C. trachomatis DNA, the detection system showed strong orange fluorescence emission peaks at 606 nm. In the presence of C. trachomatis DNA, the conformation of DNA probe changed after hybridizing with C. trachomatis DNA. Then, the amount of hairpin DNA-CuNCs was reduced and resulted in low fluorescence emission. C. trachomatis DNA displayed a significant inhibitory effect on the synthesis of fluorescent hairpin DNA-CuNCs due to the competition between C. trachomatis DNA and the specific hairpin DNA. Under the optimal experimental conditions, different concentrations of C. trachomatis were tested and the results showed a good linear relationship in the range of 50 nM to 950 nM. Moreover, the detection limit was 18.5 nM and this detection method possessed good selectivity. Finally, the fluorescent biosensor had been successfully applied to the detection of C. trachomatis target sequence in HeLa cell lysate, providing a new strategy for the detection of C. trachomatis.