A novel sandwich assay with molecular beacon as report probe for nucleic acids detection on one-dimensional microfluidic beads array |
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Authors: | Zuo Xinbing Yang Xiaohai Wang Kemin Tan Weihong Wen Jianhui |
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Institution: | State Key Laboratory of Chemo/Biosensing & Chemometrics, Biomedical Engineering Center, College of Chemistry & Chemical Engineering, Hunan University, Engineering Research Center for Bio-Nanotechnology of Hunan Province, Changsha 410082, China |
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Abstract: | A novel sandwich assay with molecular beacons as report probes has been developed and integrated into one-dimensional microfluidic beads array (1-D chip) to pursue a label-free and elution-free detection of DNA/mRNA targets. In contrast with the immobilized molecular beacons, this sandwich assay can offer lower fluorescence background and correspondingly higher sensitivity. Furthermore, this sandwich assay on 1-D chip operating in conjunction with molecular beacon technique allows multiple targets detection without the need of laborious and time-consuming elution, which makes the experiment process simple, easy to handle, and reproducible results. In the experiment, the synthesized DNA targets with different concentrations were detected with a detection limit of ∼0.05 nM. Moreover, the mRNA expression changes in A549 cells before and after anticancer drug 5-flouorouracil treatments were detected and the results were validated by the conventional RT-PCR method. |
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Keywords: | Molecular beacon Sandwich assay Microfluidic One-dimensional beads array |
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