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A sandwich plasmonic coupled surface enhanced Raman spectroscopy (SERS) tape is proposed prepared by peeling the chemical printed silver nanocorals (AgNCs) from Cu sheet with adhesive tape, which can sample targets from food surface and sandwich them between substrates and Cu sheet for SERS detection. The solid-to-solid transformation method for fabricating SERS tapes can effectively avoid the weakening of tape stickiness during the preparation process. The sandwich plasmonic coupled structure of AgNC substrate, targets, and Cu sheet display excellent SERS activity (EF = 1.62 × 107) for sensitive determination of analytes. In addition, due to the high heat conductivity of Cu sheet, the thermal effect of laser irradiation during SERS detection cannot damage the AgNC tapes, which ensures the reproducibility of subsequent quantification. The sandwich plasmonic coupled SERS tape is demonstrated to quantify malachite green (MG) and methyl parathion (MP) with good linear coefficients (> 0.98) by two typical calibration plots under different concentration ranges. The limit of detection (LOD) of the method is 0.17 ng/cm2 and 0.48 μg/cm2 (S/N = 3) for MG and MP. This method can realize the quantitative determination of MP and MG on the surface of fruits and fish scale with recoveries of 93–113%. The satisfactory detection results demonstrate the proposed sandwich plasmonic coupled AgNC tape can be successfully applied to SERS-based point-of-care testing (POCT) for pesticide residue determination, which will provide a new path for designing and constructing SERS tapes. [ABSTRACT FROM AUTHOR]

The low detection sensitivity of lateral-flow immunochromatography assay (LFIA) based on spherical gold nanoparticle (AuNP) limits its wide applications. In the present study, AuNP dimers with strong plasma scattering and robust signal output were synthesized via the Ag ion soldering (AIS) strategy and used as labeled probes in LFIA to boost the sensitivity without any extra operation process and equipment. The established LFIA exhibited high sensitivity with a limit of detection (LOD) of 2.0 × 102 TCID50/mL for PEDV, which provides 50 times higher sensitivity than commercial LFIA based on spherical colloidal gold. In addition, the AuNP dimer–based LFIA showed strong specificity, good reproducibility, high stability, and good accordance to reverse transcription polymer chain reaction (RT-PCR) when detecting 109 clinical samples. Thus, the AuNP dimers is a promising probe for LFIA and the developed AuNP dimer–based LFIA is suitable for the rapid detection of PEDV in the field. [ABSTRACT FROM AUTHOR]

Background: Approximately 1–2% of non-small cell lung cancer (NSCLC) patients harbor RET (rearranged during transfection) fusions. The oncogenic RET fusions could lead to constitutive kinase activation and oncogenesis. Methods: 1746 Chinese NSCLC patients were analyzed in this study. Tumor tissues were collected, and were formalin fixed, paraffin-embedded (FFPE) and archived. Peripheral blood (PB) samples were also collected from each patient as control. In addition, we selected 17 of them for cfDNA NGS testing and 14 tumor samples for immunohistochemistry testing using PD-L1 rabbit monoclonal antibody, clones 28-8 (Abcam, Cambridge, UK). Results: Of the 1746 NSCLC cases, RET rearrangements were identified in 25 cases (1.43%) with locally advanced or metastatic NSCLC, of which 20 (80%) were female. We found that 14 out of 25 patients had an KIF5B-RET fusion, with KIF5B exon15-RET exon12, KIF5B exon23-RET exon12, and KIF5B exon24-RET exon11 detected in 14, 3, and 1 patients, respectively. We also identified one novel RET fusion partner PLCE1 and 4 intergenic-breakpoint fusions. Conclusion: In this study, using the hybrid capture based next generation sequencing (NGS) techniques, we revealed the genomic profiling for the patients with RET fusion-positive NSCLC. To the best of our knowledge, this is the first study that exhibited the detailed breakpoints of Chinese NSCLC patients with RET rearrangement, and we found a novel new partner PLCE1. The results provided genomic information for patients with RET fusion which is significant for personalized clinical management in the era of precision medicine. [ABSTRACT FROM AUTHOR]

A novel fluorescence aptasensor based on PCN-223 as an efficient quencher was developed to sensitively detect prostate-specific antigen (PSA). The 5-carboxytetramethylrhodamine (TAMRA)-labeled PSA aptamer was adsorbed on PCN-223 by π-π stacking and hydrogen-bonding interactions, which contributed to fluorescence quenching because of the photoinduced electron transfer from TAMRA to PCN-223. In addition, the amount of quenched fluorescence of the PSA-binding aptamer complex-PCN-223 was lower than that of TAMRA aptamer-PCN-223 without PSA (at excitation/emission peaks of 545/582 nm), which can be explained by the fact that the PSA-binding aptamer complexes contributed to the separation of the aptamer from PCN-223. ∆F value of fluorescence intensities for TAMRA aptamer-PCN-223 with and without PSA showed a good linear relationship with PSA concentration over a range of 0.1 to 24 ng mL–1, with a detection limit of 0.05 ng mL–1. Compared with three metal–organic frameworks (MOFs) of UiO-66-NH2, ZIF-67, and Ni3(HITP)2 as quenchers, PCN-223 as a Zr-MOF exhibited the highest ∆F value for PSA detection. The advantage of PCN-223 could be attributed to its carboxyl, benzene, and porphyrin groups, the large specific surface area and good biocompatibility. This proposed aptasensor can be successfully used to detect PSA in sera of prostate cancer patients. The PSA detection results of this aptasensor were consistent with those which were obtained from hospital by Archtecti2000sr automatic chemiluminescence immunoanalyzer. The proposed aptasensor has potential clinical detection application. [ABSTRACT FROM AUTHOR]

The sexual strain of the planarian Schmidtea mediterranea, indigenous to Tunisia and several Mediterranean islands, is a hermaphrodite1,2. Here we isolate individual chromosomes and use sequencing, Hi-C3,4 and linkage mapping to assemble a chromosome-scale genome reference. The linkage map reveals an extremely low rate of recombination on chromosome 1. We confirm suppression of recombination on chromosome 1 by genotyping individual sperm cells and oocytes. We show that previously identified genomic regions that maintain heterozygosity even after prolonged inbreeding make up essentially all of chromosome 1. Genome sequencing of individuals isolated in the wild indicates that this phenomenon has evolved specifically in populations from Sardinia and Corsica. We find that most known master regulators5–13 of the reproductive system are located on chromosome 1. We used RNA interference14,15 to knock down a gene with haplotype-biased expression, which led to the formation of a more pronounced female mating organ. On the basis of these observations, we propose that chromosome 1 is a sex-primed autosome primed for evolution into a sex chromosome.Assembly and analysis of the Schmidtea mediterranea genome indicate that this planarian’s chromosome 1 may be evolving into a sex chromosome. [ABSTRACT FROM AUTHOR]

MicroRNAs can serve as biomarkers for many cancers, so it is significant to develop simple and sensitive strategies for microRNAs detection. Photoelectrochemical (PEC) detection has the advantages of simple equipment and high sensitivity. But in conventional PEC DNA sensors, tedious immobilization procedures of photoactive materials and capture probes on electrode surfaces are inevitable. To overcome those limitations, a homogeneous PEC biosensor based on target-responsive hydrogels has been developed (miRNA-155 has been chosen as a model target). PEC signal molecules (TiO2 nanoparticles, TiO2 NPs) were embedded in DNA hydrogels formed by hyaluronic acid sodium salt, amine-modified DNA double strands, and polyethylenimine rich in amine groups. In the presence of the target, DNA double strands in hydrogel were nicked by endonuclease and TiO2 NPs were released to the supernate and a high PEC response was obtained when collecting the supernate for PEC test, while almost no TiO2 NPs released in the absence of the target. Thanks to the exonuclease III and nicking endonuclease Nb.BbvCI-assisted cascaded amplification strategy, the proposed biosensor exhibits high sensitivity toward miRNA-155 with a low detection limit of 0.41 fM and a wide linear range from 1.0 fM to 100 pM. Since this method circumvents tedious electrode modification procedures, the proposed technique exhibits the advantages of simplicity and good reproducibility. Moreover, the prepared hydrogels have outstanding storage stability, so that they can be prepared in advance and shorten detection time. This biosensing platform provides a versatile strategy for the construction of homogeneous PEC biosensors for the detection of diverse targets. [ABSTRACT FROM AUTHOR]

A novel approach for ultrasensitive ochratoxin A (OTA) detection is reported based on dark field microscope-based single nanoparticle identification coupled with a statistical analysis method. OTA aptamers were firstly hybridized with a single-stranded DNA (DNA1) to form an identification probe (DNA1-Apt). The aptamers separate from DNA1 in the presence of OTA and are released from the identification probe. Then, another single-stranded DNA (DNA2) hybridizes with DNA1 and result in the aggregation of gold nanoparticles (AuNPs). Therefore, the presence of AuNP aggregates is the evidence of the presence of OTA, while AuNP aggregates can be easily identified together with the monomers under dark field microscopic inspection. On the other hand, by counting the aggregation rate (the number of AuNP aggregates versus the number of AuNP monomers) with a statistical analysis method, OTA could be quantitatively detected. The detection range for OTA was 0.1 pg/mL ~ 30 ng/mL and the limit of detection was 0.1 pg/mL. The proposed sensor has comparative detection performance to sensors utilizing a number of signal amplification procedures, with the additional advantages of simplicity and high efficiency. The sensor can also be adopted for other target detection simply by replacing the identification probes. [ABSTRACT FROM AUTHOR]

A fluorescence resonance energy transfer (FRET)-based in situ fluorescence signal amplification strategy is described for the determination of tyrosinase (TYR). In this assay, a dual-templated copper nanocluster (CuNCs) stabilized by bovine serum albumin (BSA) and glycylglycine (Gly-Gly) was used as an energy donor. Metyrosine was employed as a TYR substrate because its enzyme catalytic product (methyldopa) was able to function as a monomer molecule to form fluorescent polymethyldopa (PMeDP) with the assistance of BSA/Gly-Gly CuNCs. In this process, PMeDP can combine with BSA/Gly-Gly CuNCs without extra modification and then acts as an energy receptor, which leads to a remarkable FRET from BSA/Gly-Gly CuNCs to PMeDP. Interestingly, the fluorescence intensity of PMeDP was strengthened greatly in the FRET-based sensor compared to the separate excitation, which provided good sensitivity for TYR sensing. Illuminated under a UV light source, the fluorescence signal change is observed from dark violet to bright green. Therefore, the present sensing system affords a reliable ratiometric assay for TYR determination. Also, the ratio of fluorescence intensity between PMeDP (λem at 505 nm, F505) and BSA/Gly-Gly CuNCs (λem at 415 nm, F415) was used for quantitative determination of TYR. The sensing system was easily operated in aqueous media with an exciting detection limit of 44.0 U L−1. This sensing strategy has been applied to the screening of inhibitors. [ABSTRACT FROM AUTHOR]

Bisphenol A (BPA), a typical endocrine disruptor, is widely used as a key monomer in the packaging industry. Residual monomer can transfer from the package material to the food and thereby pose a risk to the health of the consumer, so determination of BPA migration is highly important for food safety control. In this study, a simple but sensitive electrochemiluminescence (ECL) biosensor, which combines the characteristics of high selectivity of an aptamer and high sensitivity of ECL, has been developed to detect BPA from package materials. The aptamer was immobilized on a gold electrode surface through Au-S interaction. The aptamer was then hybridized with complementary DNA (CDNA) to form double-stranded DNA (dsDNA). Ru(phen) can intercalate into the grooves of dsDNA and acts as an ECL indicator; high ECL intensity can therefore be detected from the electrode surface. In the presence of BPA, which can competitively bind with the aptamer owing to their high affinity, Ru(phen) is released from the electrode surface and the ECL of the system is decreased. The decreasing ECL signal has a linear relationship with BPA in the range of 0.1-100 pM with a detection limit of 0.076 pM. The developed biosensor has been applied to detect migration of BPA from different categories of canned drink with satisfactory results. [ABSTRACT FROM AUTHOR]

Telomerase is one of the most common markers of human malignant tumors, such as uterine, stomach, esophageal, breast, colorectal, laryngeal squamous cell, thyroid, bladder, and so on. It is necessary to develop some sensitive but convenient detection methods for telomerase activity determination. In this study, a label-free and ultrasensitive electrochemiluminescence (ECL) biosensor has been fabricated to detect the activity of telomerase extracted from HeLa cells. Thiolated telomerase substrate (TS) primer was immobilized on the gold electrode surface through gold-sulfur (Au-S) interaction and then elongated by telomerase specifically. Then, it was hybridized with complementary DNA to form double-stranded DNA (dsDNA) fragments on the electrode surface, and Ru(phen) has been intercalated into the dsDNA grooves to act as the ECL probe. The enhanced ECL intensity has a linear relationship with the number of HeLa cells in the range of 5∼5000 and with a detection limit of 2 HeLa cells. The proposed ECL biosensor has high specificity to telomerase in the presence of common interferents. The relative standard deviations (RSDs) were <5 % at 100 HeLa cells. The proposed method provides a convenient approach for telomerase-related cancer screening or diagnosis. [ABSTRACT FROM AUTHOR]

A rapid method is described for the preparation of a highly uniform and sensitive SERS substrate by an improved 'drop-and-dry' method. Gold nanobipyramids (Au NBPs) were prepared inside the nanoholes (nanowalls) of anodic aluminum oxide (AAO) templates with a typically 5-μm nanohole depth. The SERS substrate can be prepared by this method within 40 s and on large scale. The SERS signals obtained with this Au NBPs-AAO substrate is stronger by four-orders of magnitude compared to conventional a silicon wafer substrate. The SERS signal for dopamine (DA; measured at 1311 cm−1) is found to be enhanced by a factor of 2.2 × 108. The response to DA extends from 10 nM to 0.1 mM, and the limit of detection is 6.5 nM (at S/N = 3). The assay was applied to the determination of DA in spiked human serum. Graphical abstract Schematic presentation of a highly active and uniform 3-dimensional substrate composed of gold nanobipyramids and anodic aluminum oxide (Au NBP/AAO). It was used for on-spot sensing of dopamine. [ABSTRACT FROM AUTHOR]

A copper(II)-based two-dimensional metal-organic framework with nanosheet structure (CuBDC NS) that possesses peroxidase (POx) mimicking activity was prepared. In the presence of hydrogen peroxide, the system catalyses the oxidation of terephthalic acid to a blue-fluorescent product (excitation = 315 nm; emission = 425 nm). Pyrophosphate has a very strong affinity for Cu2+ ion and blocks the POx-mimicking activity of the CuBDC NS. If, however, inorganic pyrophosphatase is present, the POx mimicking activity is gradually restored because pyrophosphate is hydrolyzed. The findings were used to design a method for the determination of the activity of inorganic pyrophosphatase by fluorometry. Fluorescence increases linearly in the 1-50 mU·mL−1 inorganic pyrophosphatase activity range. The limit of detection is 0.6 mU·mL−1 (S/N = 3).A copper(II)-based two-dimensional metal-organic framework (CuBDC NS) is described that possesses POx-mimicking activity. Inorganic pyrophosphate (PPi) was hydrolyzed to phosphate in the presence of inorganic pyrophosphatase (PPase). Hence, it cannot coordinate with Cu2+ in CuBDC NS, its structure was well-conserved to catalyses the oxidation of terephthalic acid (H2BDC) to produce a blue fluorescent product (oxBDC) in the presence of hydrogen peroxide (H2O2). [ABSTRACT FROM AUTHOR]

A surface-enhanced Raman scattering (SERS) substrate with good flexibility and high water absorbing capacity is reported. It consists of a calcium alginate sponge incorporating gold nanoparticles. These are in close contact with the sponge without the need for amino or sulfhydryl modification. The substrate is capable of detecting the dyes crystal violet (CV) and malachite green (MG) in water directly and rapidly by immersing it into the liquid sample. Preconcentration and separation are not required. The dyes absorbed on the sponge can be detected without drying and thus the whole analytical process can be completed within 3 min. The results show that the lowest detectable concentrations are 0.1 and 0.25 μg⋅L−1 for CV and MG, respectively. This is lower than the minimum required performance limits set by the European Commission and the US EPA. Moreover, MG and CV can be simultaneously detected in liquid samples due to their different SERS bands (at 1216 and 1534 cm−1, respectively). It should be noted that the molecular structures of MG and CV are very similar. Therefore, the method has a large potential for determination of several analytes simultaneously even in complex sample metrics.Schematic presentation of the fabrication of a sodium alginate sponge loaded with gold nanoparticles. Gold nanoparticles together with gel-like alginate were freeze-dried to form the sponge. The sponge was cross-linked by CaCl2 solution and then it was freeze-dried again. [ABSTRACT FROM AUTHOR]

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 × 102 cfu·mL−1 in a short time (within 15 min), and the working range is 1 × 102 to 1 × 106 cfu·mL−1, which is superior over the detection performance of conventional test strip using AuNRs modified by centrifugation.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. [ABSTRACT FROM AUTHOR]

A portable, affordable and simple detector is requested in a 'Point-of-Care-Testing' (POCT) system. In this study, we exploited the potentialities of Differential Pressure Gauge (DPG) to the orientation of POCT technology. Alpha fetoprotein (AFP) was chosen as a model analyte that could specifically recognized by its antigen, and a tiny outfits equipped with a DPG was employed as the signal readout. Pt/SiO2 nanospheres were synthesized and modified with the detection antibody. In the presence of target, a sandwich of immunocomplex specifically formed and the Pt/SiO2 had been modified on the capture antibody. Which then can be dissolved to release plenty of Pt and the suspensions were transferred into a closed vial filled with appropriated amount of hydrogen peroxide. Subsequently, hydrogen peroxide was decomposed to produce oxygen, resulting in the enhancement of pressure in the closed vial and which can be detected by DPG easily. Under the optimized conditions, the read out signal from DPG had a direct relationship with AFP concentrations in the range of 10~200 ng/mL, and the detection limit was as low as 3.4 ng/mL. The proposed portable sensor had been successfully applied to detect AFP in serum samples with satisfactory results. This strategy holds a great promising in biological analysis as its convenient operations, reliable results and flexible apparatus. [ABSTRACT FROM AUTHOR]

In this work, 3,3′,5,5′-tetramethylbenzidine(II) (TMB2+), derived from H2O2-horseradish peroxidase (HRP)-3,3′,5,5′-tetramethylbenzidine (H2O2-HRP-TMB) reaction system, was used to etch AuNRs to generate different colors of solution. Many enzyme reactions are involved in the production of H2O2 (e.g., glucose can react with the dissolved oxygen in the presence of glucose oxidase (GOx) to produce H2O2). Given this information, a simple visual biosensor was developed in this study, with glucose as the example target. The detection range of the proposed system varied with the experimental conditions, such as the concentration of GOx and HRP, and enzymatic reaction time. Under the optimized conditions, the longitudinal shift of localized surface plasmon resonances (LSPR) had a linear correlation with the glucose concentration in the range of 0.1~1.0 mM. Meanwhile, the solution displayed a specific color in response to the glucose concentration, thus enabling the visual quantitative detection of glucose at a glance. Compared with the traditional monochromic colorimetry, this multicolor glucose sensor generates various vivid colors, which can be easily distinguished by naked eyes without any sophisticated instrument. Notably, the proposed method has been successfully applied to detect glucose in serum samples with satisfied results. [ABSTRACT FROM AUTHOR]

Surface enhanced spectroscopy such as surface enhanced Raman spectrum (SERS) and surface enhanced fluorescence have been investigated extensively in the past two decades. Herein, we present experimental evidence to demonstrate the existence of a new surface enhanced spectroscopy, namely, surface enhanced electrochemiluminescence (SEECL). Our investigation indicates that the electrochemiluminescence (ECL) response of the Ru(bpy)32+-tri-n-propylamine (TPrA) system could be significantly enhanced when the working electrode is modified with gold nanoparticle-SiO2 core-shell nanocomposites (AuNP@SiO2). It is worth noting that comparing with a working electrode modified with pure SiO2 nanoparticles, the electrochemical responses of the two electrodes were quite similar, but the ECL signal of the AuNP@SiO2 modified electrode was ~5 times higher than that of the SiO2 nanoparticles modified electrode. Thus we infer that the localized surface plasmon resonance (LSPR) of the AuNPs could be a major contribution to the ECL enhancement. Our investigations also demonstrate that the ECL enhancement is closely related to the thickness of the SiO2 layer. As much as 10 times ECL enhancement (comparing with the ECL intensity of bare electrode) is observed under the optimal conditions. The possible mechanism of the SEECL phenomenon is also discussed. [ABSTRACT FROM AUTHOR]

Surface enhanced spectroscopy such as surface enhanced Raman spectrum (SERS) and surface enhanced fluorescence have been investigated extensively in the past two decades. Herein, we present experimental evidence to demonstrate the existence of a new surface enhanced spectroscopy, namely, surface enhanced electrochemiluminescence (SEECL). Our investigation indicates that the electrochemiluminescence (ECL) response of the Ru(bpy)32+-tri-n-propylamine (TPrA) system could be significantly enhanced when the working electrode is modified with gold nanoparticle-SiO2 core-shell nanocomposites (AuNP@SiO2). It is worth noting that comparing with a working electrode modified with pure SiO2 nanoparticles, the electrochemical responses of the two electrodes were quite similar, but the ECL signal of the AuNP@SiO2 modified electrode was ~5 times higher than that of the SiO2 nanoparticles modified electrode. Thus we infer that the localized surface plasmon resonance (LSPR) of the AuNPs could be a major contribution to the ECL enhancement. Our investigations also demonstrate that the ECL enhancement is closely related to the thickness of the SiO2 layer. As much as 10 times ECL enhancement (comparing with the ECL intensity of bare electrode) is observed under the optimal conditions. The possible mechanism of the SEECL phenomenon is also discussed. [ABSTRACT FROM AUTHOR]

Loss of heterozygosity through inbreeding or mitotic errors leads to reductions in progeny survival and fertility. Loss of heterozygosity is particularly exacerbated in geographically isolated populations, which are prone to inbreeding depression and faster rates of extinction. The regenerative capacities of the hermaphroditic biotype of the planarian Schmidtea mediterranea allowed us to perform a systematic genetic test of Mendelian segregation and study the loss of heterozygosity in the Spiralian superclade in general and planarians in particular. We discovered that ~300 Mb (~37.5%) of the genome retains heterozygosity even after ten generations of inbreeding, and show that these chromosomal regions have low diversity and recombination rates in wild populations. Our genetic and genomic analyses establish S. mediterranea as a genetically tractable system. The research also opens the door to study the evolutionary basis of non-Mendelian mechanisms, the adaptive advantages of chromosome structural heterozygotes and their potential relationship to the robust regenerative capacities of planarians.