Strategy For Simultaneous Detection Research Articles

Utilization of Schiff base ligands to chelate Ag+ for highly specific and sensitive Hg2+/Ag+ immunoassay

In this study, a planar benzothiazole Schiff base derivative ZX1 was designed and synthesized to selectively precipitate silver ions, in conjunction with the immunoassay detection of Hg2+ ions. The silver chelation process was highly effective in the pH range of 7 to 10, with a silver ion precipitation ratio of 99.9 %. The coordination behavior of ZX1 with Ag+ was studied using a series of spectroscopic measurements, topography analysis and DFT calculation. The shortest bond distances were 2.251 and 2.254 Å between nitrogen atom of thiazole and Ag+. Weak interactions were analyzed using the interaction region indicator (IRI) method and VMD. The immunoassay was developed through an anti-Hg2+/Ag+ monoclonal antibody, which showed similar sensitivities for Ag+ and Hg2+. The selectivity and anti-interference ability of the Schiff base ligands were tested for some hard metal ions and sevreal anions. With co-existence of interference ion Ag+, Hg2+ was detected in rice matrix solution with 77 %∼123.5 % recovery by addition of ZX1 as the masking reagent and 90.3 %∼115 % for wheat matrix. This research provides a promising immunoassay strategy for simultaneous detection of silver ions and mercury ions in food matrix by utilization of Schiff base ligands. This novel Schiff base ligand was then used as the pre-treatment reagent to remove silver ions within 5 min for detection of Hg2+ or detection of both Ag+ and Hg2+ based on a novel bi-specific heavy metal antibody.

Eu-doped carbon dots-MOF based turn-on fluorescent probe for trace Hg2+ and Pb2+ and construction of the simultaneous detection model

Multiple trace heavy metal pollutants present a serious threat to the aquatic environment, and their coexistence also affected the detection efficiency. This work prepared Eu doped metal organic framework (MOF) complexed with S-containing carbon dots (CD(s)) exhibited dual-emission fluorescence. Its chelation-enhanced fluorescence with heavy metals replaced the original agglomerative fluorescence quenching of pure CD(s), which was utilized to construct a “turn on” fluorescence sensing probe. Based on the XPS, DFT and fluorescence lifetime results, the N, S functional groups on the surface of the materials could bind specifically to Hg2+ and Pb2+ with enhanced sensitivity. The limit of detection reached 76.67 nM for Hg2+ and 5.12 nM for Pb2+ within 2 min detection time. Furthermore, due to the different response mechanism of the two signal peaks to Hg2+ and Pb2+, a three-dimensional data model was developed for simultaneous detection in the co-existence system. The model had been successfully applied to actual Hg2+-Pb2+ containing water samples with recoveries ranged from 93.15 ∼ 105.08 %. The results provided a new strategy for simultaneous detection of trace environmental pollutants.

Rapid detection and differentiation of chlortetracycline and tetracycline by N,P-Ti3C2 QDs

Ti3C2 QDs with good water solubility and photoluminescence properties was prepared by nitrogen and phosphorus doping (N,P-Ti3C2 QDs). The probe has a fluorescence emission peak at 420 nm and emits bright blue fluorescence, which can distinguish and detect two antibiotics with similar structures, chlortetracycline (CTC) and tetracycline (TC). The fluorescence enhancement effect can be realized by the interaction with CTC. The linear range is 2–22 μM, and the detection limit is as low as 0.53 μM. And the change of fluorescent color from bright blue to bright purple can be observed with the naked eye. At the same time, with the addition of TC, the fluorescence of N,P-Ti3C2 QDs can be quenched by static quenching effect, and the fluorescence color changes from bright blue to yellowish green. The detection range is 4.1–20.5 μM, and the detection limit is as low as 0.16 μM.Therefore, on this basis, a strategy for simultaneous detection and recognition of CTC and TC is proposed. The method has been successfully applied to the determination of CTC and TC in milk samples. This paper provides a new detection method for the identification of tetracycline antibiotics and taps the application potential of Ti3C2 QDs in food detection.

Magnetic photoelectrochemical sensor array utilizing addressing sensing strategy for simultaneous detection of amyloid-β 42 and microtubule-associated protein tau

The accurate diagnosis of diseases can be improved by detecting multiple biomarkers simultaneously. This study presents the development of a magnetic photoelectrochemical (PEC) immunosensor array for the simultaneous detection of amyloid-β 42 (Aβ) and microtubule-associated protein (Tau), which are markers for neurodegenerative disorders. A metal-organic framework (MOF) derivative, Fe2O3@FeS2 magnetic composites with exceptional photoelectric and ferromagnetic properties was synthesized while preserving the original structure and advantages. Thus, the immunoassembly process of the sensor can be carried out in homogeneous solution and recovered by magnetic separation. For simultaneous detection, a chip is divided into multiple independent sensing sites, which have the same preparation and detection environment, allowing for the implementation of a self-calibration method. The sensor array demonstrates considerable detection ranges of 0.01–100 ng mL−1 for Aβ and 0.05–100 ng mL−1 for Tau, with low detection limits of 2.1 pg mL−1 for Aβ and 7.9 pg mL−1 for Tau. The PEC sensor array proposed in this study exhibits exceptional stability, selectivity, and reproducibility, providing a new method for detecting multiple markers.

Dual-functional metal-organic frameworks-based hydrogel micromotor for uranium detection and removal

Self-propelled micro/nanomotors have attracted great attention for environmental remediation, however, their use for radioactive waste detection and removal has not been addressed. Engineered micromotors that are able to combine fast detection and highly adsorptive capability are promising tools for radioactive waste management but remain challenging. Herein, we design self-propelled micromotors based on zeolite imidazolate framework (ZIF-8)-hydrogel composites via inverse emulsion polymerization and show their potential for efficient uranium detection and removal. The incorporation of magnetic ferroferric oxide nanoparticles enables the magnetic recycling and actuation of the single micromotors as well as formation of swarms of worm-like or tank-treading structure. Benefited from the enhanced motion, the micromotors show fast and high-capacity uranium adsorption (747.3 mg g−1), as well as fast uranium detection based on fluorescence quenching. DFT calculation confirms the strong binding between carboxyl groups and uranyl ions. The combination of poly(acrylic acid-co-acrylamide) with ZIF-8 greatly enhances the fluorescence of the micromotor, facilitating the high-resolution fluorescence detection. A low detection limit of 250 ppb is reached by the micromotors. Such self-propelled micromotors provide a new strategy for the design of smart materials in remediation of radioactive wastewater.

Magnetically-assisted electrochemical immunoplatform for simultaneous detection of active and total prostate-specific antigen based on proteolytic reaction and sandwich affinity analysis

Simultaneous detection of active and inactive proteases is clinically meaningful for improving diagnostic specificity. In this work, we reported an electrochemical method for simultaneous immunoassays of active and total proteases. Magnetic beads (MBs) were used as the solid supports for immobilization of capture antibodies and enrichment of targets. For the detection of active protease, the proteolytic-reaction-based analysis was carried out by the generation of Cu2+-binding peptide, in which a label-free peptide was used as the proteolytic substrate. The redox potential of the resulting peptide-Cu2+ complex was intrinsically distinguished from that of free Cu2+, thus allowing the “signal-on” detection of active protease. For the immunoassay of total protease in a sandwich-like format, electroactive metal-organic frameworks (Cu-MOFs) were used as the signal tags. The captured Cu-MOFs could directly produce a well-defined electrochemical signal from the reduction of Cu2+ ions. The analytical performances of the immunoplatform were evaluated by determining the model analytes of free and total prostate-specific antigen (fPSA and tPSA) in buffer and serum. The detection limits were found to be 0.3 pM for fPSA and 2 pM for tPSA. This work proposed a new strategy for simultaneous detection of active and total proteases, which should be evaluable for clinical diagnosis and treatment of protease-relative diseases.

FDM-assisted opposite two-way OA-CEAS system employing four lasers for simultaneous multi-species detection.

An FDM-assisted opposite two-way OA-CEAS system is reported in this paper. Compared with the traditional OA-CEAS system with one-way transmission configuration, the new system has two main advantages. One of the advantages is that four lasers can be employed for simultaneous measurements of multiple species in this system. Another advantage is the combination of the silver-coated concave spherical mirror and the narrow bandpass filter employed to realize the opposite two-way transmission in the optical cavity which can also serve as a re-injection mirror and optical enhancement gotten for free in the system. The performance of the system is demonstrated by simultaneous measurements of CO, CO2, C2H4, and CH4. This work highlights a new strategy for simultaneous detection by using four lasers in a single optical integrated cavity, which can improve the utilization rate of the optical cavity and reduce the cost for multiple gas species sensing.

A spatial-potential-resolved electrochemiluminescence biosensor for simultaneous detection of BRCA1 and BRCA2 based on a novel self-luminescent metal–organic framework

Conventional potential-resolved electrochemiluminescence (ECL) biosensors often require the participation of multiple co-reactants, which greatly limits their practical application. To solve the problem, a new kind of self-luminescent copper-based metal–organic framework (Cu-MOF) was synthesized for the first time, Cu-MOF can produce superior positive potential ECL without co-reactants. A novel spatial-potential-resolved electrochemiluminescence biosensor based on Cu-MOF and CdTe@CdS quantum dots (QDs) was constructed for simultaneous detection of BRCA1 and BRCA2. To achieve positive and negative potential luminescence at the same time, Cu-MOF and CdTe@CdS QDs were integrated into different locations of ITO electrode to achieve spatial-potential-resolved ECL only using one co-reactant, which had not been previously reported to my knowledge. Cu-MOF was placed in channel a as the signal luminophor, and Au@Ag NPs were used to quench the positive potential ECL of Cu-MOF by resonance energy transfer (RET) for detection of target BRCA1. Similarly, CdTe@CdS QDs as negative potential ECL material were introduced into channel b for detection of BRCA2. This work not only developed a novel self-luminescent ECL material, but also proposed an ingenious sensing strategy for simultaneous detection of multi-target, which has a wide application prospect in the field of biochemical analysis.

Potential-resolved electrochemiluminescence biosensor for simultaneous determination of multiplex miRNA

The multi-target simultaneous detection strategy based on potential-resolved electrochemiluminescence (ECL) has still been a research hotspot in analytical science, but the limited selection of ECL luminophores hinders the development of this field. Herein, polyethyleneimine functionalized perylene derivatives (PTC-PEI) and luminol functionalized gold nanoparticles (Lu–Au NPs) possessed significantly resolved emission potentials as ECL luminophore. The ternary ECL system was constructed with MoS2 nanoflowers and K2S2O8 as the coreaction accelerator and coreactant respectively, which significantly improved the cathode ECL emission of PTC-PEI. Simultaneously, the anode coreaction accelerator ZnO nanoflowers could promote the anode coreactant dissolved O2 reduction, and extremely enhanced the anode ECL emission of Lu–Au NPs. The proposed strategy addressed the major technical challenge of cross interference and competition of the coreactants for dual-biomarker detection, thus enabling accurate detection of miRNA-205 and miRNA-21 from 10 fM to 100 nM, with detection limits of 2.57 and 1.15 fM, respectively. In general, this work achieved a single-step synchronous detection of dual biomarkers, providing a new idea for the ECL detection of multiple biomarkers, and having potential value in the clinical diagnosis.

Two-color lateral flow nucleic acid assay combined with double-tailed recombinase polymerase amplification for simultaneous detection of chicken and duck adulteration in mutton

We propose a visual strategy for simultaneous detection of chicken and duck ingredients in mutton by integration of a double tailed multiplex recombinase polymerase amplification (mRPA) assay with a multicolor nucleic acid hybridization lateral flow strip (mNAH-LFS) assay. Two colored gold nanoparticles (AuNPs), including gold nanospheres (red) and gold nanoflowers (blue), were synthesized and functionalized with the corresponding nucleic acid probe to target each analyte. The RPA product was analyzed by this two-colored mNAH-LFS assay which includes a red T1 line for chicken adulteration and a blue T2 line for duck adulteration. In this RPA reaction, special Spacer9 and tail nucleotide sequence-tagged primers (Tag+Spacer9+F-primer, Tag+Spacer9+R-primer) were used to obtain double-stranded amplicons with specific wobbled sequences that can directly bind with AuNPs-labeled probes (AuNSs-cT1 hybridized with T1, AuNFs-cT4 hybridized with T4) and solid-phase probes (biotin-cT2 hybridized with T2, biotin-cT3 hybridized with T3) on the mNAH-LFS producing a red or/and cyan color on the T lines. The detection limits of this mRPA-mNAH-LFS assay for chicken and duck were 0.01% and 0.01%, respectively. Finally, this mRPA-mNAH-LFS assay was successfully used to authenticate 59 commercial meat products. In conclusion, this mRPA-mNAH-LFS is a sensitive, accurate, and cost-effective approach for meat quality assurance and traceability.

Major benznidazole metabolites in patients treated for Chagas disease: Mass spectrometry-based identification, structural analysis and detoxification pathways

Benznidazole is the drug of choice for the treatment of Chagas disease, but its metabolism in humans is unclear. Here, we identified and characterized the major benznidazole metabolites and their biosynthetic mechanisms in humans by analyzing the ionic profiles of urine samples from patients and untreated donors through reversed-phase UHPLC-ESI-QTOF-MS and UHPLC-ESI-QqLIT-MS. A strategy for simultaneous detection and fragmentation of characteristic positive and negative ions was employed using information-dependent acquisitions (IDA). Selected precursor ions, neutral losses, and MS3 experiments complemented the study. A total of six phase-I and ten phase-II metabolites were identified and structurally characterized in urine of benznidazole-treated patients. Based on creatinine-corrected ion intensities, nitroreduction to amino-benznidazole (M1) and its subsequent N-glucuronidation to M5 were the main metabolic pathways, followed by imidazole-ring cleavage, oxidations, and cysteine conjugations. This extensive exploration of benznidazole metabolites revealed potentially toxic structures in the form of glucuronides and glutathione derivatives, which may be associated with recurrent treatment adverse events; this possibility warrants further exploration in future clinical trials. Incorporation of this knowledge of the benznidazole metabolic profile into clinical pharmacology trials could lead to improved treatments, facilitate the study of possible drug-drug interactions, and even mitigation of adverse drug reactions.

A molecularly imprinted fiber array solid-phase microextraction strategy for simultaneous detection of multiple estrogens.

A kind of selective enrichment material based on a homemade molecularly imprinted polymer (MIP) fiber array with high adsorption capacity was developed for the accurate analysis of estrogens in food samples. Here, the MIP with 17β-estradiol as the template was obtained by in situ polymerization. The chemical composition, morphologies, surface area, and pore size of the polymer were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and Brunauer-Emmett-Teller theory. The extraction time, desorption solvent, desorption time, ionic strength, and the pH of solution were investigated to ascertain the best extraction conditions. Under the optimal extraction conditions, three fiber coatings of 17β-estradiol MIP and commercial polyacrylate (PA) were bound to a homemade handle to assemble the fiber array, respectively. The findings showed that the three-fiber array of the MIP significantly improved the extraction capacity by 145 times compared to PA. The MIP fiber array showed high adsorption capacity for the template molecule 17β-estradiol and its structural analogues estrone, bisphenol F, bisphenol B, and bisphenol A, with enrichment factors of 99.60-133.16. A molecularly imprinted polymer solid-phase microextraction fiber array (MIP-SPME fiber array) coupled with high-performance liquid chromatography-diode array detection was used for the analysis and detection of the five estrogens in milk and yogurt samples. Satisfactory recoveries were achieved ranging from 74.75-119.41% with <9.42% relative standard deviations. The developed method for the simultaneous determination of trace estrogens in food samples exhibited a limit of detection of 0.33 μg L-1. The MIP-SPME fiber array provided an available strategy for improving the selectivity and adsorption capacity of SPME for trace target component analysis in complex matrices and increasing the sensitivity of the analytical method.

Ultrasensitive dual-enhanced sandwich strategy for simultaneous detection of Escherichia coli and Staphylococcus aureus based on optimized aptamers-functionalized magnetic capture probes and graphene oxide-Au nanostars SERS tags

In this work, a novel surface-enhanced Raman scattering (SERS) sandwich strategy biosensing platform has been established for simultaneously detecting Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). Fe3O4@SiO2-Au nanocomposites (NCs) with varying amounts of Au nanocrystals were prepared, and the effect of interparticle gaps on SERS activity was studied by finite-difference time-domain (FDTD) method. The optimal magnetic SERS-active substrates (FS-A5) were functionalized with the specific aptamers to act as capture probes. Meanwhile, graphene oxide-Au nanostars (GO-Au NSs) decorated with Raman reporters and aptamers were used as SERS tags. The loading density of Au NSs on GO was tuned to change the number of SERS active sites. In this proposal, E. coli and S. aureus were first captured by capture probes and then bound with SERS tags to form a sandwich-like structure, which caused enhanced electromagnetic field because of the dual enhancement strategy. Under optimal conditions, SERS platform could detect E. coli and S. aureus simultaneously, and the detection limit was as low as 10 cfu/mL. Our sandwich assay-based dual-enhanced SERS platform provides a new idea for simultaneously detecting multiple pathogens with high selectivity and sensitivity, and thus will have more hopeful prospects in the field of food safety.

Antibody-free photoelectrochemical strategy for simultaneous detection of methylated RNA, METTL3/METTL14 protein and MazF protein based on enhanced photoactivity of MoSe2–BiOI nanocomposite

Taking advantages of the catalytic activity of METTL3/METTL14 protein towards adenine methylation in RNA sequence and the specific digestion activity of MazF protein towards unmethylated RNA sequence containing ACA bases, a novel photoelectrochemical biosensor was constructed for simultaneous detection of RNA methylation, METTL3/METTL14 protein and MazF protein. MoSe2–BiOI nanocomposite was prepared and considered as photoactive material, catalytic hairpin assembly strategy and in situ generation of electron donors catalyzed by polyaspartic acid-loaded alkaline phosphatase technique were employed as signal amplification. Under the optimum conditions, the detection ranges of methylated RNA, METTL3/METTL14 protein and MazF protein were 0.001–50 nM, 0.001–25 ng/μL, and 0.001–10 U/mL, respectively. The corresponding detection limits were 0.46 pM, 0.51 pg/μL and 0.42 U/μL with S/N = 3. In addition, the effect of drugs and composite pollutants on the activities of MazF proteins was assessed, proving the applicability of the developed method in the field of drug screening for MazF-related diseases. Moreover, the effects of pollutants on the activity of METTL3/METTL14 were also preliminarily explored, providing new information on pathogenic mechanism of pollutants.

A simple and smart AND-gate DNA nanoprobe for correlated enzymes tracking and cell-selective imaging

Accurate cancer diagnosis and effective drug therapy entail sensitive and dynamic monitoring of intracellular key enzymes, since their expression level is closely related to disease progression. Simultaneous monitoring of correlated enzymes is promising to help unveiling mystery of cytobiological events during tumor progression and drug response, while is challenged by lacking of a robust and simple simultaneous detection strategy. In order to construct a simple and smart strategy which is complex design-avoided and doesn't need other auxiliary enzyme, here we develop an AND-gate strategy for simultaneously monitoring correlated enzymes which both are upregulated in cancer cells (telomerase and apurinic/apyrimidinic endonuclease 1). An innovative AND-gate DNA nanoprobe has been designed to avoid mutual interference and background noise, guaranteeing an enhanced fluorescent signal output upon catalyzation of dual enzymes. This AND-gate strategy achieves sensitive detection of two enzymes in an individual manner in test tube, through which the diagnostic potential of bladder cancer has been validated by telomerase detection in clinical urine sample. The AND-gate strategy enables specific intracellular imaging of dual enzymes in different cancer cell lines. Importantly, in contrast to traditional single-targeting strategies, AND-gate imaging of dual enzymes significantly improves cancer cell selectivity. Moreover, this strategy dynamically monitors enzymatic activity changes during chemoresistance induced by chemotherapeutic treatment. This simple and smart strategy has foreseeable prospect in the fields of disease diagnosis, drug prognosis evaluation, and precise fluorescence-guided surgery.

Freeze-synthesized bio-barcode immunoprobe based multiplex fluorescence immunosensor for simultaneous determination of four nitrofuran metabolites

Here we have reported a simple and sensitive bio-barcode immunosensor for simultaneous detection of 3-amino-2-oxazolidinone (AOZ), 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), 1-aminohydantoin (AHD), and semicarbazide (SEM) in aquatic products. According to freeze–thaw strategy, four fluorophores (FAM, HEX, ROX, Cy5) labeled single-stranded DNA (ssDNA) were conjugated onto the surface of gold nanoparticles (AuNPs) with corresponding four nitrofuran metabolites monoclonal antibodies (mAbs) for forming four bio-barcode fluorescence immunoprobes. The fluorescence of immunoprobes was quenched by AuNPs. In test progress, the ssDNA with fluorophores were released by adding the dithiothreitol (DTT) and the fluorescence recovered. The immunosensor exhibited sensitive and specific detection of nitrofuran metabolites from 0.05 to 28 μg/L. The limit of detection (LOD) was 0.01, 0.02, 0.02, and 0.05 μg/L for AOZ, AMOZ, AHD, and SEM, respectively. The recoveries of four nitrofuran metabolites in spiked aquatic products have been confirmed by UPLC-MS/MS. The bio-barcode based multiplex immunosensor provides a promising strategy for simultaneous detection of multiple targets.

Multiplexed flow cytometric approach for detection of anti-SARS-CoV-2 IgG, IgM and IgA using beads covalently coupled to the nucleocapsid protein.

Flow cytometry has emerged as a promising technique for detection of SARS‐CoV‐2 antibodies. In this study, we developed an innovative strategy for simultaneous detection of immunoglobulin G (IgG), IgM and IgA. The SARS‐CoV‐2 nucleocapsid protein was covalently bound to functional beads surface applying sulpho‐SMCC chemistry. BUV395 anti‐IgG, BB515 anti‐IgM, biotinylated anti‐IgA1/IgA2 and BV421 streptavidin were used as fluorophore conjugated secondary antibodies. Serum and antibodies reaction conditions were optimized for each antibody isotype detection and a multiplexed detection assay was developed. This new cell‐free assay efficiently discriminate COVID‐19 negative and positive samples. The simultaneous detection of IgG, IgM and IgA showed a sensitivity of 88·5–96·2% and specificity of 100%. This novel strategy opens a new avenue for flow cytometry‐based diagnosis.

Cell-based flow cytometry assay for simultaneous detection of multiple autoantibodies in a single serum sample

Accurate serologic evaluation of autoantibodies in patients with autoimmune diseases is critical. In the present study, we established a live cell-based assay for simultaneous detection of multiple autoantibodies in a single serum sample. Autoantibody seropositivity was determined by 3-color flow cytometry using live Chinese hamster ovary cells transiently expressing a target protein of interest fused to enhanced green fluorescent protein and labeled with Alexa Fluor 647 and Hoechst 33342. As a representative example, we applied the strategy for simultaneous detection of 2 recently established biomarkers for central nervous system autoimmune inflammatory demyelinating disorders, anti-aquaporin-4 autoantibody and anti-myelin oligodendrocyte glycoprotein autoantibody, in a single serum sample. This analysis revealed the coexistence of these 2 autoantibodies. We demonstrated that this assay can simultaneously detect 3 different autoantibodies. We propose a quadrant gating strategy of flow cytometry contour plots to clearly distinguish seropositive sera from seronegative sera regardless of the extent of the background signal level or the autoantibody titer. This novel and practical method using a combination of fluorescent proteins and fluorochromes to simultaneously detect multiple autoantibodies improves the efficiency of evaluating serum samples, and therefore provides significant benefits to both the patient and the healthcare professionals performing autoantibody testing.

Targeted RNA sequencing for upfront analysis of actionable driver alterations in non-small cell lung cancer

ObjectivesTargeted RNA-based Next-Generation Sequencing (tRNA-seq) is increasingly being used in molecular diagnostics for gene fusion detection in non-small cell lung cancer (NSCLC). However, few data support its clinical application for the detection of single nucleotide variants (SNVs) and small insertions/deletions. In this study, we evaluated the performance of tRNA-seq using Archer FusionPlex for simultaneous detection of actionable gene fusions, splice variants, SNVs and indels in formalin-fixed, paraffin-embedded NSCLC tissue. Materials and methodsA total of 126 NSCLC samples, including 20 validation samples and 106 diagnostic cases, were analyzed by targeted DNA-based Next-Generation Sequencing (tDNA-seq) followed by tRNA-seq. ResultsAll 28 SNVs and indels in the validation set, and 34 out of 35 mutations in the diagnostic set were identified by tRNA-seq. The only mutation undetected by tRNA-seq, ERBB2 p.(Ser310Tyr), was not included in the current Archer panel design. tRNA-seq revealed one additional BRAF p.(Val600Glu) mutation not found by tDNA-seq. SNVs and indels were correctly called by the vendor supplied software, except for ERBB2 duplication p.(Tyr772_A775dup) which was only detected by an additional in-house developed bio-informatics pipeline. Variant allelic frequency (VAF) values were generally higher at the expression level compared to the genomic level (range 6–96% for tRNA-seq versus 6–61% for tDNA-seq) and low VAF mutations in DNA (6–8% VAF) were all confirmed by tRNA-seq. Finally, tRNA-seq additionally identified a driver fusion or splice variant in 10 diagnostic NSCLC samples including one MET exon 14 skipping variant not detected by tDNA-seq. ConclusionOur results demonstrate that tRNA-seq can be implemented in a diagnostic setting as an efficient strategy for simultaneous detection of actionable gene fusions, splice variants, SNVs and indels in NSCLC provided that adequate RNA-seq analysis tools are available, especially for the detection of indels. This approach allows upfront identification of currently recommended targetable molecular alterations in NSCLC samples.

A low-cost high-entropy porous CrO/CrN/C biosensor for highly sensitive simultaneous detection of dopamine and uric acid

A low-cost high-entropy porous CrO/CrN/C biosensor was fast fabricated by mechanical milling strategy for simultaneous detection of dopamine (DA) and uric acid (UA). Biosensor carrier derived from the natural waste jasmine flowers (JF) could be coated by DA and then coordinated with Cr3+ with the aid of high-energy mechanochemistry, which fabricate porous Cr-JFC/DAC electrode with highly dispersed CrO/CrN sites. The formed large amount of high active sites significantly enhanced the biosensor’s response signal and resolution for the detection of DA and UA. Moreover, the high surface (950.1 m2/g), enriched interface and conductive carbon carrier efficiently improved the electrical signal transfer of biosensor. The Cr-JFC/DAC2 modified electrode was used for the simultaneous determination of DA and UA in the range of 0.05 ∼ 4 μM with a limit of detection (LOD) 10.65 and 17.54 nM (S/N = 3) respectively. The LOD were 1 ∼ 2 orders of magnitude less than reported state-of-the-art non-precious metal sensors, and even lower than those of most precious metal sensors. The analytical performance of the Cr-JFC/DAC2-modified electrodes has been evaluated using human serum samples with recoveries of 97.00%∼104.00%, good anti-interference and stability. Therefore, the Cr-JFC/DAC2/Nafion/GCE exhibited a great potential in the real application to simultaneously detect DA and UA.