A quantum dot-based microRNA nanosensor for point mutation assays

Abstract

We have developed a quantum dot-based microRNA nanosensor for point mutation assays using primer generation-mediated rolling circle amplification. The proposed method exhibits high sensitivity with a detection limit of as low as 50.9 aM and a large dynamic range of 7 orders of magnitude from 0.1 fM to 1 nM. Importantly, this method can be further applied to analyze the point mutation of mir-196a2 in the lung tissues of non small-cell lung cancer patients.

miRNA Optical Detection

“…The combination of QDs with single-molecule detection (e.g., nucleic acids, proteins, and small molecules) enables the development of QDbased biosensors with distinct advantages including high signal-to-noise ratios, low sample consumption, and high sensitivity [27]. In particular, QDs hold great promise as fluorescence resonance energy transfer (FRET) [28] or electrochemiluminescence resonance energy transfer (ERET) [29] donors in various biosensors for miRNA detection. Zhang and coworkers [28] developed a QD-based miRNA nanosensor for a point mutation assay with primer generation-mediated rolling circle amplification (PG-RCA), and further applied it to analyze the point mutation of mir-196a2 in real samples.…”

“…In particular, QDs hold great promise as fluorescence resonance energy transfer (FRET) [28] or electrochemiluminescence resonance energy transfer (ERET) [29] donors in various biosensors for miRNA detection. Zhang and coworkers [28] developed a QD-based miRNA nanosensor for a point mutation assay with primer generation-mediated rolling circle amplification (PG-RCA), and further applied it to analyze the point mutation of mir-196a2 in real samples. As shown in Fig.…”

Advance in quantitative single-molecule detection

“…http://engine.scichina.com/doi/10.1360/N032017-00074 图 2 蛋白激酶分析示意图 [29] (网络版彩图) 图 3 基于单个量子点的纳米传感器用于hOGG1分析的示 意图 [30] (网络版彩图) Zhang课题组 [33] 将基于FRET的单分子检测与滚环 扩增和酶切反应相结合, 用于分析非小细胞肺癌病人组 [34] (网络版彩图)…”

Coupling hybridization chain reaction with catalytic hairpin assembly enables non-enzymatic and sensitive fluorescent detection of microRNA cancer biomarkers

“…Moreover, the integration of two different signal amplification routes into one single assay can further push down the detection limit and significantly improve the sensitivity. Such integrated approaches have proved to be very useful for ultrasensitive detection of different biomarkers Zeng et al, 2014). These methods, however, require the use of nanomaterial/enzyme signal amplification labels as aforementioned, which increases the cost and complexity of the assay protocols.…”