An ultrasensitive and specific electrochemiluminescence (ECL) biosensor has been designed for the p53 DNA sequence, which is based on cascade signal amplification of nicking endonuclease assisted target recycling and hyperbranched rolling circle amplification (HRCA). First of all, biotin modified hairpin capture DNA (HP) probe was immobilized on the surface of streptavidin magnespheres paramagnetic particles (PMPs). Target DNA hybridized with the loop portion of the HP probe, therefore unfolding HP to form a double-stranded DNA (dsDNA) containing the specific nicking site of the nicking endonuclease. Then, the nicking endonuclease recognized the specific nicking site and cleaved the HP into two pieces, liberating target DNA and the complementary sequence piece for the padlock probe. The intact target DNA would initiate the next cycle of hybridization and cleavage, thereby releasing multiple complementary sequences for the padlock probes. The liberated complementary sequences hybridized with the padlock probes, subsequently inducing the HRCA reaction and generating numerous dsDNA segments. Herein, Ru(phen)3(2+) was embedded into dsDNA and worked as ECL signal reporter. The reaction products were eventually pretreated by dialysis tube with the cutoff membrane to remove the residual Ru(phen)3(2+) in the solution for the following ECL measurements. Using this cascade amplification strategy, an ultrasensitive p53 DNA sequence detection method was developed with a wide linear range from 0.05 to 100 fM and a low detection limit of 0.02 fM. Moreover, this cascade amplified ECL biosensor had specific recognition capacity for noncomplementary and single- and double-base mismatched DNA. The proposed ECL biosensor might have a great potential in biomedical research and clinic analysis.
We report a very easy and effective approach for synthesizing unique palladium-on-gold supra-nanostructure (Au@Pd-SprNS)-decorated graphene oxide (GO) nanosheets. The SprNSs comprising Au nanorods as core and a unique close-packed assembly of tiny anisotropic Pd nanoparticles (NPs) as shell were homogeneously distributed on the GO surface via electrostatic self-assembly. Compared with the traditional one-pot method for synthesis of metal NPs on GO sheets, the size and shape of core-shell Au@Pd SprNSs can be finely controlled and uniformly distributed on the GO carrier. Interestingly, this Au@Pd-SprNSs/GO nanocomposite displayed high electrocatalytic activities toward the oxidation of methanol, ethanol, and formic acid, which can be attributed to the abundance of intrinsic active sites including high density of atomic steps, ledges and kinks, Au-Pd heterojunctions and cooperative action of the two metals of the SprNSs. Additionally, uniform dispersion of the SprNSs over the GO nanosheets prevent agglomeration between the SprNSs, which is of great significance to enhance the long-term stability of catalyst. This work will introduce a highly efficient Pd-based nanoelectrocatalyst to be used in fuel cell application.
Ligand exchange on the surface of gold nanorods (AuNRs) is widely used, but conventional methods usually require multiple centrifugation cycles to completely remove cetyltrimethylammonium bromide (CTAB). This can lead to undesired aggregation of AuNRs. A dialysis-assisted protocol is described here for ligand exchange on AuNRs. Dialysis driven by a concentration gradient is shown to be a powerful tool to separate CTAB from aqueous solutions. The concentration gradient of CTAB in a dialysis bag can avoid the possible aggregation of AuNRs that can be caused by drastic environmental changes. It also supports the rate of ligand exchange on the surfaces of the AuNRs. The modified AuNRs were employed in a lateral-flow test strip immunoassay (LFTS-IAs) for the food pathogen E. coli O157:H7 in order to study of efficiency of ligand exchange. Compared to AuNRs where ligand exchange was performed via multiple centrifugation cycles, the AuNRs prepared by dialysis-assisted ligand exchange show improved conjugation to antibody and enhanced visual signals in the test line of the LFTS-IAs. A portable strip reader (absorption wavelength = 525 nm) is used to records the testing results. The sensitivity of AuNRs modified by dialysis has been achieved even as low as 1 × 10 cfu·mL in a short time (within 15 min), and the working range is 1 × 10 to 1 × 10 cfu·mL, which is superior over the detection performance of conventional test strip using AuNRs modified by centrifugation. Graphical abstract Schematic presentation of the ligand exchange of AuNRs. The AuNRs were dialysed in water to decrease the CTAB concentration. Then, 11-mercaptoundecanoic acid (MUA) replaces the CTAB capped on the surface of AuNRs. The modified AuNRs were employed in a lateral flow immunoassay for E. coli O157:H7.
The
global spread of SARS-CoV-2 virus has severely affected human
health, life, and work. Vaccine immunization is considered to be an
effective means to protect the body from infection. Therefore, timely
analysis of the antibody level is helpful to identify people with
low immune response or attenuated antibodies so as to carry out targeted
and precise vaccine booster immunization. Herein, we develop a magnetofluid-integrated
multicolor immunochip, as a sample-to-answer system in a fully enclosed
space, for visual analysis of neutralizing antibodies of SARS-CoV-2.
Generally, this chip adopts an innovative three-dimensional two-phase
system that utilizes mineral oil to block the connection between reagent
wells in the vertical direction and provides a wide interface for
rapid and nondestructive shuttle of magnetic beads during the immunoassay.
In order to obtain visualized signal output, gold nanorods with a
size-dependent color effect are used as the colorful chromogenic substrates
for evaluation of the antibody level. Using this chip, the neutralizing
antibodies were successfully detected in vaccine-immunized volunteers
with 83.3% sensitivity and 100% specificity. Furthermore, changes
in antibody levels of the same individual over time were also reflected
by the multicolor assay. Overall, benefiting from simple operation,
airtight safety, and nonrequirement of external equipment, this platform
can provide a new point-of-care testing strategy for alleviating the
shortage of medical resources and promoting epidemic control in underdeveloped
areas.
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