Rolling circle amplification based colorimetric determination of Staphylococcus aureus

文献类型： 外文期刊

第一作者： Li, Yanan

作者： Li, Yanan;Wang, Junping;Wang, Junying;Wang, Shuo

作者机构：

关键词： Rolling circle amplification (RCA); Colorimetric microplate assay; Multifunctional gold nanoparticles; Biotin-streptavidin system; Staphylococcus aureus

期刊名称：MICROCHIMICA ACTA （ 影响因子：5.833； 五年影响因子：5.357 ）

ISSN： 0026-3672

年卷期： 2020 年 187 卷 2 期

页码：

收录情况： SCI

摘要： A colorimetric microplate assay for determination of Staphylococcus aureus DNA is described. Linear padlock probes were designed to recognize target sequences. After DNA binding, the linear padlock probes were circularized by ligation and then hybridize with biotin-labeled capture probes. Biotin-labeled capture probes act as primers to initiate the RCA. The biotin-labeled RCA products hybridize with digoxin-labeled signal probes fixed on streptavidin-functionalized wells of a 96-well plate. To enhance sensitivity, an AuNP-anti-digoxigenin-POx-HRP conjugate was added to the wells and then bound to digoxin-labeled signalling probes. The oxidation of tetramethylbenzidine (TMB) by H2O2 produces a color change from colorless to blue via HRP catalysis. After the reaction was terminated, absorbance is measured at 450 nm. For target sequences of Staphylococcus aureus, the detection limit is 1.2 pM. For genomic DNA, the detection limit is 7.4 pg.mu L-1. The potential application of the method was verified by analyzing spiked food samples.

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