Perform Signal Amplification Research Articles

Electrochemical Biosensing Platform Based on Toehold-Mediated Strand Displacement Reaction and DSN Enzyme-Assisted Amplification for Two-Target Detection.

Simultaneous detection of multiple targets is of great significance for accurate disease diagnosis. Herein, based on duplex-specific nuclease (DSN) assisted signal amplification and the toehold-mediated strand displacement reaction (TSDR), we constructed an electrochemical biosensor with high sensitivity and high specificity for dual-target detection. MiRNA-141 and miRNA-133a were used as the targets, and ferrocene (Fc) and methylene blue (MB) with significant peak potential differentiation were used as the electrochemical signal probes. The elaborately designed hairpin probe H1, which was fixed on the electrode surface, could be hybridized with the target miRNA-141 to perform signal amplification by the DSN-assisted enzyme cleavage cycle; thus, miRNA-141 could be detected by Fc signal changes at 0.41 V. The hairpin H1 can also combine with the MB-labeled signal probe (SP) output from miRNA-133a-induced TSDR, and the detection of miRNA-133a can be realized according to the response signal generated by MB at -0.26 V. The two sensing lines are independent of each other, and there is no mutual interference in the detection process. Therefore, two independent detection lines could be connected in series, and the simultaneous detection of two targets can be achieved on a single electrode. This novel detection strategy provides a new way to simultaneously detect different biomarkers.

Rapid and ultrasensitive detection of ellagic acid by integrating boronate-affinity controllable-oriented imprinted magnetic nanoparticle and boronic acid-modified / polyethylene glycol-coated allochroic-graphene oxide

Herein, we integrate boronate-affinity controllable-oriented ellagic acid-imprinted magnetic nanoparticles (BCEMN) and boronic acid-modified / polyethylene glycol-coated allochroic-graphene oxide (BPAO) for specific and ultrasensitive detection of ellagic acid (EA). The novel detection method depends on a dual boronate-affinity sandwich strategy. The symmetric EA is first specifically captured by the BCEMN based on the boronate-affinity-related molecularly imprinted recognition, and then labeled with BPAO due to the boronate-affinity recognition. The BPAO, which is functionalized with 2,6-difluoro-4-formyl-phenylboronic acid, loaded with a large amount of pH-sensitive thymol phthalide and coated with hydrophilic polyethylene glycol, can specifically recognize the cis-diol of EA at physiological condition (pH 7.4) and perform signal amplification and readout through pH-triggered allochroism. Furthermore, the signal readout of allochroism is linear with the logarithm of EA concentration over the range between 0.02 ng mL−1 to 104 ng mL−1 with a good correlation coefficient (R2 = 0.993), and the limit of detection (LOD) is 2.96 pg mL−1. Finally, the BCEMN linked BPAO (BCBP) strategy has also been successfully applied in the detection of EA concentration in spiked strawberry sample with recovery and relative standard deviation in the range of 84.9%–110.2% and 3.9%–6.1%, respectively.

Visual detection of hepatocellular carcinoma cells with cell imprinted substrate and pH-sensitive allochroic-graphene oxide

Herein, we integrate cell-imprinted substrate (CIS) and allochroic-graphene oxide (AGO) for specific visualization sorting of hepatocellular carcinoma cells. The state-of-the-art-of detection method relies on the enzyme linked immunosorbent assay (ELISA)-like sandwich strategy with hierarchical recognition. The target tumor cells are first selectively captured by the CIS based on cell imprinted recognition, and then specifically labeled with AGO by boronate affinity recognition between boronic acid on AGO and cis-diols on the surface of target cells. The selectively recognition of CIS for target template cells is verified by cell function experiments. It is also worth mentioning that the AGO can specifically recognize target tumor cells under physiological pH, and then perform signal amplification and output through pH-triggered allochroism. The CIS linked AGO for cell assay (CIS-AGO-CA) is successfully used for visualization detection of human hepatocarcinoma HLE cells from hepatocyte suspension. When the hepatocyte suspension is spiked with 1.0 × 105 cells, the recoveries of CIS-AGO-CA are 80.67 ± 4.33% for target HLE cells, and only 12.00 ± 1.00% for non-target Hep3B cells. It is worth emphasizing that the CIS-AGO-CA process is antibody-free. Therefore, this novel ELISA-like sandwich strategy is high specificity, cost-efficient and easy-to-use, and exhibits great prospect in the visualization sorting of tumor subpopulation.

Highly sensitive chemiluminescent point mutation detection by circular strand-displacement amplification reaction

Single nucleotide polymorphism (SNP) genotyping is attracting extensive attentions owing to its direct connections with human diseases including cancers. Here, we have developed a highly sensitive chemiluminescence biosensor based on circular strand-displacement amplification and the separation by magnetic beads reducing the background signal for point mutation detection at room temperature. This method took advantage of both the T4 DNA ligase recognizing single-base mismatch with high selectivity and the strand-displacement reaction of polymerase to perform signal amplification. The detection limit of this method was 1.3 × 10 −16 M, which showed better sensitivity than that of most of those reported detection methods of SNP. Additionally, the magnetic beads as carrier of immobility was not only to reduce the background signal, but also may have potential apply in high through-put screening of SNP detection in human genome.

Catalysis in Reaction Networks

We define catalytic networks as chemical reaction networks with an essentially catalytic reaction pathway: one which is "on" in the presence of certain catalysts and "off" in their absence. We show that examples of catalytic networks include synthetic DNA molecular circuits that have been shown to perform signal amplification and molecular logic. Recall that a critical siphon is a subset of the species in a chemical reaction network whose absence is forward invariant and stoichiometrically compatible with a positive point. Our main theorem is that all weakly-reversible networks with critical siphons are catalytic. Consequently, we obtain new proofs for the persistence of atomic event-systems of Adleman etal., and normal networks of Gnacadja. We define autocatalytic networks, and conjecture that a weakly-reversible reaction network has critical siphons if and only if it is autocatalytic.

Chemical signal amplification in two-dimensional paper networks

Two-dimensional paper networks (2DPNs) hold great potential for extending the utility of paper-based chemical and biochemical diagnostics at a cost and ease-of-use that are comparable to conventional lateral flow strips. 2DPNs enable the automated sequential delivery of multiple reagents to a detection region with a single user activation step, and therefore have the potential to extend the processing capabilities of inexpensive paper-based assays with comparable ease of use to conventional lateral flow tests. In this communication, we used a simple 3 inlet 2DPN to perform signal amplification of a colloidal gold label using a gold enhancement solution, thus demonstrating the capability of 2DPNs to perform processes that can improve limits of detection.

Preliminary results of an in-beam PET prototype for proton therapy

Proton therapy can overcome the limitations of conventional radiotherapy due to the more selective energy deposition in depth and to the increased biological effectiveness. Verification of the delivered dose is desirable, but the complete stopping of the protons in patient prevents the application of electronic portal imaging methods that are used in conventional radiotherapy During proton therapy β + emitters like 11C, 15O, 10C are generated in irradiated tissues by nuclear reactions. The measurement of the spatial distribution of this activity, immediately after patient irradiation, can lead to information on the effective delivered dose. First, results of a feasibility study of an in-beam PET for proton therapy are reported. The prototype is based on two planar heads with an active area of about 5×5 cm 2. Each head is made up of a position sensitive photomultiplier coupled to a square matrix of same size of LYSO scintillating crystals (2×2×18 mm 3 pixel dimensions). Four signals from each head are acquired through a dedicated electronic board that performs signal amplification and digitization. A 3D reconstruction of the activity distribution is calculated using an expectation maximization algorithm. To characterize the PET prototype, the detection efficiency and the spatial resolution were measured using a point-like radioactive source. The validation of the prototype was performed using 62 MeV protons at the CATANA beam line of INFN LNS and PMMA phantoms. Using the full energy proton beam and various range shifters, a good correlation between the position of the activity distal edge and the thickness of the beam range shifter was found along the axial direction.