MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University , Guangzhou 510631, China.
ACS Appl Mater Interfaces. 2014 Jul 9;6(13):9988-96. doi: 10.1021/am503230h. Epub 2014 Jun 13.
Graphene oxide (GO) and quantum dots (QDs), as burgeoning types of nanomaterials, have gained tremendous interest in the biosensor field. In this work, we designed a novel multivariate testing strategy that depends on the fluorescence resonance energy transfer (FRET) effect between quantum dots (QDs) and graphene oxide (GO). It integrates the QD-GO FRET principle and QD probes with different emission peaks into a platform, aims at multiplex gene detection of a human infectious and highly pathogenic pathogen, Listeria monocytogenes (L. monocytogenes). With the development of a multiplex linear-after-the-exponential (LATE) polymerase chain reaction (PCR) system, the single-stranded DNA (ssDNA) products of hlyA genes and iap genes are obtained by simultaneous amplification of the target genes. Then with the hybridization of ssDNA products and QD probes, simultaneous homogeneous detection of two gene amplification products can be achieved by using GO as a fluorescence switch and monitoring the relevant emissions excited by a single light source. Distinguishable signals corresponding to target genes are obtained. With this developed approach, genomic DNA from L. monocytogenes can be detected as low as 100 fg/μL. Moreover, this platform has a good dynamic range from 10(2) to 10(6) fg/μL. It is indicated that this platform has potential to be a reliable complement for rapid gene detection technologies and is capable of reducing the false-negative and false-dismissal probabilities in routine tests.
氧化石墨烯（GO）和量子点（QDs）作为新兴的纳米材料，在生物传感器领域引起了极大的兴趣。在这项工作中，我们设计了一种新颖的多元测试策略，该策略依赖于量子点（QDs）和氧化石墨烯（GO）之间的荧光共振能量转移（FRET）效应。它将 QD-GO FRET 原理和具有不同发射峰的 QD 探针集成到一个平台中，旨在对人类感染性和高致病性病原体李斯特菌（Listeria monocytogenes，L. monocytogenes）的多个基因进行检测。通过建立一个多元线性滞后（LATE）聚合酶链反应（PCR）系统，目标基因的同时扩增得到 hlyA 基因和 iap 基因的单链 DNA（ssDNA）产物。然后，通过 ssDNA 产物与 QD 探针的杂交，利用 GO 作为荧光开关，监测单光源激发的相关发射，可以实现两种基因扩增产物的同时均相检测。得到与目标基因相对应的可区分信号。利用该方法，最低可检测到 100 fg/μL 的李斯特菌基因组 DNA。此外，该平台的动态范围为 10(2)到 10(6) fg/μL。这表明该平台具有成为快速基因检测技术可靠补充的潜力，能够降低常规检测中的假阴性和假阴性概率。
