The outbreak of the pandemic caused by the severe acute respiratory syndrome
coronavirus-2 (SARS-CoV-2) calls for an urgent unmet need for developing a facial and
cost-effective detection method. The requirement of well-trained personnel and
sophisticated instrument of current primary mean (reverse transcription polymerase chain
reaction, RT-PCR) may hinder the practical application worldwide. In this regard, a
reverse transcription recombinase polymerase amplification (RT-RPA) coupled with
CRISPR-Cas12a colorimetric assay is proposed for the SARS-CoV-2 detection. The
methodology we have described herein utilizes DNA-modified gold nanoparticles (AuNPs) as
a universal colorimetric readout and can specifically target ORF1ab and N regions of the
SARS-CoV-2 genome. After the virus genome is amplified through RT-RPA, the resulting
abundant dsDNA will bind and activate Cas12a. Under trans-cleavage degradation, the
capped DNA substrate will be hydrolyzed gradually from AuNPs, demonstrating a change in
the surface plasmon resonance (SPR), which can be facially monitored by UV–vis
absorbance spectroscopy and naked eye observation. The high amplification efficiency
from RT-RPA and Cas12a trans-cleavage process bring the sensitivity of our method to 1
copy of viral genome sequence per test. Notably, under the dual variations inspecting
from the isothermal amplification and Cas12a activation process, the false positive
events from other beta coronavirus members can be effectively avoided and thus
significantly improve the specificity. Furthermore, the reliability of this colorimetric
assay is validated by standard clinical samples from the hospital laboratory department.
Through integration of the inherently high sensitivity and specificity from an
RPA-coupled Cas12a system with the intrinsic simplicity of AuNP-based colorimetric
assay, our method increases the practical testing availability of SARS-CoV-2.
Noble metal nanoparticles are promising catalysts in electrochemical reactions, while understanding the relationship between the structure and reactivity of the particles is important to achieve higher efficiency of electrocatalysis, and promote the development of single-molecule electrochemistry. Electrogenerated chemiluminescence (ECL) was employed to image the catalytic oxidation of luminophore at single Au, Pt, and Au-Pt Janus nanoparticles. Compared to the monometal nanoparticles, the Janus particle structure exhibited enhanced ECL intensity and stability, indicating better catalytic efficiency. On the basis of the experimental results and digital simulation, it was concluded that a concentration difference arose at the asymmetric bimetallic interface according to different heterogeneous electron-transfer rate constants at Au and Pt. The fluid slip around the Janus particle enhanced local redox reactions and protected the particle surface from passivation.
The development of highly sensitive and selective uranyl ion (UO22+) probes has attracted significant attention owing to the threat to human health caused by high toxicity, radioactivity, and long half‐life. Herein, the development of aggregation‐induced emission (AIE) active polymer dots (Pdots) is described for an accurate UO22+ monitoring using a portable electrochemiluminescence (ECL) system. An AIE‐active polymer containing tetraphenylethene and boron ketoiminate moieties is prepared into Pdots and modified with ssDNA to capture UO22+, which can amplify the ECL signal of the Pdots through a resonance energy transfer mechanism. This probe provides an ultralow detection limit of 10.6 pm/2.5 ppt, which is at least two orders of magnitude lower than the known UO22+ luminescent probes. Only UO22+ can provide an obvious ECL enhancement among the various metal ions, indicating the excellent selectivity of this probe. Furthermore, a portable ECL analyzer is designed to realize UO22+ measurements in the wild. The anodic ECL mechanism of UO22+ is discovered and ECL technology is first applied in monitoring radioactive substances. This study provides a novel strategy for the development of accurate UO22+ probes and a practical UO22+ monitoring method, indicating its potential application in the environmental and energy fields.
In this work, a simple and label-free electrochemical biosensor is developed for microRNA (miRNA) detection on the basis of an arched probe mediated isothermal exponential amplification reaction (EXPAR). The arched probe assembled on the electrode surface consists of two strands that are partially complementary to each other at both ends. The target can hybridize with the complementary sequence of the arched structure, leading to the cleavage of the probe. The strand fixed on the surface of the electrode self-assembles, in the presence of hemin, to G-quadruplex unit, yielding electrochemical signals. The other strand liberated into the solution triggers the EXPAR to recycle and regenerate targets. This method exhibits ultrahigh sensitivity toward miRNA with detection limits of 5.36 fM and a detection range of 3 orders of magnitude. The biosensor is capable of discriminating a single-nucleotide difference between concomitant miRNA and performs well in analyzing crude extractions from cancer cell lines.
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