Multifunctional Photoelectrochemical Biosensor Based on ZnIn2S4/ZnS QDs@Au-Ag-Reversed Photocurrent of Cu-Metal-Organic Framework Coupled with CRISPR/Cas-12a-Shearing for Assay of Dual Targets.

False positives and negatives in bioanalytical assays remain a persistent problem. Herein, a multifunctional photoelectrochemical (PEC) biosensor based on ZnIn2S4 (ZIS)/ZnS quantum dots (QDs)@Au-Ag-reversed photocurrent of Cu-metal-organic framework (MOF) coupled with CRISPR/Cas-12a-shearing was innovatively developed for assay of dual targets. First, Cu-MOF as a good PEC material shows cathodic photocurrent. Then, numerous ZIS/ZnS QDs were assembled to the Au-Ag nanoparticles (NPs) to prepare a stable and highly amplified signal probe, which can just match the energy level of Cu-MOFs and realized the polarity-reversed photocurrent of Cu-MOF for the first time. As the empty-core nanostructure of Au-Ag NPs has a high specific surface area and low material density, the bimetallic nanocrystal can much increase the reaction rate and improve the redox efficiency. When target CEA-produced cDNA opened the hairpin DNA (HP1 DNA) on the electrode, the ZIS/ZnS QDs@Au-Ag signal probe was conjugated to the electrode via DNA hybridization, achieving a significantly reversed PEC current for CEA detection. Moreover, the specific binding of kanamycin/aptamer generated the acDNA (activator), which can activate the trans-cleavage activity of the CRISPR-CAS12a system on ssDNA, so the signal probe was sheared and caused the obvious decrease of PEC signal for kanamycin detection. The newly developed ZIS/ZnS QDs@Au-Ag NPs displayed excellent PEC properties and reversed photocurrent to MOF and were combined with the unique CRISPR-Cas12a system to achieve sensitive detection of dual targets, which can open a new polarity-reversed PEC sensing platform for rapid and accurate analysis of multiple targets and can effectively avoid false positives results in clinical testing.

Photoelectrochemical biosensing platform based on in situ generated ultrathin covalent organic framework film and AgInS2 QDs for dual target detection of HIV and CEA.

In this work, a new photoelectrochemical (PEC) biosensing platform based on an ordered two-dimensional (2D) ultrathin covalent organic framework (COF) film and AgInS2 quantum dots (QDs) has been developed to enable dual-target detection of HIV and CEA. The porous COF film was firstly in situ generated on ITO, displaying super-stable and intense photocurrent with excellent repeatability. Moreover, an effective PEC quenching probe was specifically designed by loading large number of AgInS2 QDs on Au nanoparticles (NPs). After target HIV-induced cyclic amplification process to generate abundant DNA S0, the Au NPs-AgInS2 QDs probe was binded to the COF film through DNA hybridization, enabling PEC signal of the COF film to turn "off" for ultra-sensitive detection of HIV. Furthermore, when CEA as the second target specifically binded to its aptamer, the Au NPs-AgInS2 QDs quenching probe was released, achieving PEC signal "on" of the T-DA COF film for ultra-sensitive detection of CEA. This work opened a unique 2-D COF film-based PEC biosensing platform with excellent signal for rapid detection of dual targets, which can effectively avoid false positives and negatives and shows promising application for early prevention and detection of cancer diseases.

Dual-wavelength electrochemiluminescence biosensor based on a multifunctional Zr MOFs@PEI@AuAg nanocomposite with intramolecular self-enhancing effect for simultaneous detection of dual microRNAs.

Rapid parallel detection of multi-targets has always been an exploration aim in electrochemiluminescence (ECL) assays. Herein, a multifunctional nanocomposite of Zr metal-organic frameworks (MOFs) @PEI@AuAg nanoclusters (NCs) with intense and stable dual-wavelength ECL was synthesized for the first time, and used to construct a new ECL biosensor for rapid simultaneous detection of dual targets. Notably, the novel ECL emitter Zr MOFs with high-performance was not only integrated with a co-reactant polyethyleneimine (PEI) to form a unique intramolecular self-enhancing structure, but also loaded a large number of another ECL emitter AuAg NCs, furthermore, AuAg NCs with superior electron transfer property can much enhance the electrical conductivity of the composites, thus achieving the goal of "killing three birds with one stone". Moreover, a unique stable and rigid three-dimensional DNA tetrahedron (TDN) structure was connected with two quenching probes BHQ1 and BHQ3 and immobilized on the composites-modified electrode, so ECL emission of the nanocomposites at two wavelengths of 535 nm and 644 nm were both quenched by resonance energy transfer (RET). In the presence of target miRNAs, the efficient DNA cycling double-amplification processes were performed by using exonuclease (T7 Exo) combined with DNA Walker, thus both quenching groups were separated to restore the ECL at two wavelengths, achieving simultaneous and rapid ECL detection of two miRNAs. Therefore, this present work not only opens a unique nanocomplex with dual wavelength ECL and self-enhancing performance, but also develops a highly sensitive ECL biosensor with promising value for rapid multi-target analysis in clinical fields.

ß-Cyclodextrin-functionalized graphene and metal-organic framework composites for ultrasensitive electrochemical detection of chloramphenicol.

Novel ß-cyclodextrin (ß-CD)@functionalized graphene (G)/Cu-1,3,5-benzenetricarboxylic acid (BTC) composites were in situ prepared using ß-CD functionalized graphene and Cu-BTC, and a new electrochemical sensor for sensitive detection of chloramphenicol (CAP) was developed based on the composites. A series of investigations on the ß-CD@G/Cu-BTC composite material were conducted. The ß-CD functionalized graphene solution has an excellent and stable dispersion effect. The composite was further combined with metal-organic frameworks (MOFs) to overcome the disadvantages of a single material, displaying excellent conductivity and a synergistic catalytic effect on the detection of chloramphenicol; so an electrochemical sensor for CAP detection is developed. An actual sample was also detected using the proposed sensor.

Electrochemiluminescence resonance energy transfer biosensing platform between g-C3N4 nanosheet and Ru-SiO2@FA for dual-wavelength ratiometric detection of SARS-CoV-2 RdRp gene.

Rational detection of syndrome coronavirus 2 (SARS-CoV-2) is crucial to prevention, control, and treatment of disease. Herein, a dual-wavelength ratiometric electrochemiluminescence (ECL) biosensor based on resonance energy transfer (RET) between g-C3N4 nanosheets and Ru-SiO2@folic acid (FA) nanomaterials was designed to realize ultrasensitive detection of SARS-CoV-2 virus (RdRp gene). Firstly, the unique g-C3N4 nanosheets displayed very intense and stable ECL at 460 nm, then the triple helix DNA was stably and vertically bound to g-C3N4 on electrode by high binding affinity between ssDNA and g-C3N4. Meanwhile, trace amounts of target genes were converted to a large number of output by three-dimensional (3D) DNA walker multiple amplification, and the output bridged a multifunctional probe Ru-SiO2@FA to electrode. Ru-SiO2@FA not only showed high ECL at 620 nm, but also effectively quenched g-C3N4 ECL. As a result, ECL decreased at 460 nm and increased at 620 nm, which was used to design a rational ECL biosensor for detection of SARS gene. The results show that the biosensor has excellent detection sensitivity for RdRp gene with a dynamic detection range of 1 fM to 10 nM and a limit of detection (LOD) of 0.18 fM. The dual-wavelength ratio ECL biosensor has inestimable value and application prospects in the fields of biosensing and clinical diagnosis.

Versatile Photoelectrochemical Biosensing for Hg2+ and Aflatoxin B1 Based on Enhanced Photocurrent of AgInS2 Quantum Dot-DNA Nanowires Sensitizing NPC-ZnO Nanopolyhedra.

Eliminating false positives or negatives in analysis has been a challenge. Herein, a phenomenon of polarity-switching photocurrent of AgInS2 quantum dot (QD)-DNA nanowires reversing nitrogen-doped porous carbon-ZnO (NPC-ZnO) nanopolyhedra was found for the first time, and a versatile photoelectrochemical (PEC) biosensor with a reversed signal was innovatively proposed for dual-target detection. NPC-ZnO is a photoactive material with excellent PEC properties, while AgInS2 QDs as a photosensitive material match NPC-ZnO in the energy level, which not only promotes the transfer of photogenerated carriers but also switches the direction of PEC current. Furthermore, in order to prevent spontaneous agglomeration of AgInS2 (AIS) QDs and improve its utilization rate, a new multiple-branched DNA nanowire was specially designed to assemble AgInS2 QDs for constructing amplified signal probes, which not only greatly increased the load of AgInS2 QDs but also further enhanced the photoelectric signal. When the target Hg2+-induced cyclic amplification process generated abundant RDNA, the DNA nanowire signal probe with plenty of QDs was linked to the NPC-ZnO/electrode by RDNA, generating greatly amplified polarity-reversed photocurrent for signal "ON" detection of Hg2+. After specific binding of the target (aflatoxin B1, AFB1) to its aptamer, the signal probes of AIS QD-DNA nanowires were released, realizing signal "OFF" assay of AFB1. Thus, the proposed new PEC biosensor provides a versatile method for detection of dual targets and also effectively avoids both false positive and negative phenomena in the assay process, which has great practical application potential in both environmental and food analysis.