Rapid, on-site quantitative determination of mycotoxins in grains using a multiple time-resolved fluorescent microsphere immunochromatographic test strip.

The label probe plays a crucial role in enhancing the sensitivity of lateral flow immunoassays. However, conventional fluorescent microspheres (FMs) have limitations due to their short fluorescence lifetime, susceptibility to background fluorescence interference, and inability to facilitate multi-component detection. In this study, carboxylate-modified Eu(III)-chelate-doped polystyrene nanobeads were employed as label probes to construct a multiple time-resolved fluorescent microsphere-based immunochromatographic test strip (TRFM-ICTS). This novel TRFM-ICTS facilitated rapid on-site quantitative detection of three mycotoxins in grains: Aflatoxin B1 (AFB1), Zearalenone (ZEN), and Deoxynivalenol (DON). The limit of detection (LOD) for AFB1, ZEN, and DON were found to be 0.03 ng/g, 0.11 ng/g, and 0.81 ng/g, respectively. Furthermore, the TRFM-ICTS demonstrated a wide detection range for AFB1 (0.05-8.1 ng/g), ZEN (0.125-25 ng/g), and DON (1.0-234 ng/g), while maintaining excellent selectivity. Notably, the test strip exhibited remarkable stability, retaining its detection capability even after storage at 4 °C for over one year. Importantly, the detection of these mycotoxins relied solely on simple manual operations, and with a portable reader, on-site detection could be accomplished within 20 min. This TRFM-ICTS presents a promising solution for sensitive on-site mycotoxin detection, suitable for practical application in various settings due to its sensitivity, accuracy, simplicity, and portability.

Rational design of peptide-based fluorescent probe for sequential recognitions of Cu(II) ions and glyphosate: Smartphone, test strip, real sample and living cells applications.

A new peptide-based fluorescent probe named DMDH with easy-to-synthesize, excellent stability, good water solubility and large Stokes shift (225 nm) was synthesized for highly selective sequential detections of copper ions (Cu2+) and glyphosate (Glyp). DMDH demonstrated great detection performance towards Cu2+via strong fluorescence quenching, and forming non-fluorescence DMDH-Cu2+ ensemble. As a new promising cascade probe, the fluorescence of DMDH-Cu2+ ensemble was significantly recovered based on displacement approach after glyphosate was added. Interestingly, the limit of detections (LODs) for Cu2+ and glyphosate were 40.6 nM and 10.6 nM, respectively, which were far lower than those recommended by the WHO guidelines for drinking water. More importantly, DMDH was utilized to evaluate Cu2+ and glyphosate content in three real water samples, demonstrating that its effectiveness in water quality monitoring. Additionally, it is worth noting that DMDH was also applied to analyze Cu2+ and glyphosate in living cells in view of significant cells permeability and low cytotoxicity. Moreover, DMDH soaked in filter paper was used to create qualitative test strips and visually identify Cu2+ and glyphosate through significant color changes. Furthermore, smartphone RGB color recognition provided a new method for semi-quantitative testing of Cu2+ and glyphosate in the absence of expensive instruments.

Development of Colloidal Gold Test Strips for the Detection of Oxamyl Residues in Tobacco.

In this study, monoclonal antibodies against oxamyl were prepared, and colloidal gold immunochromatography was used to design a rapid test strip product for the detection of oxamyl in tobacco with high specificity, accuracy and stability without cross-reactivity to commonly used tobacco fungicides based on the optimization of conditions such as pH value of diluent, diluent dosage, concentration of antibody marker, type of confining solution and complex solution. 5 The results of five samples of post-harvest ready-to-bake tobacco and first-harvest tobacco were consistent with the gas chromatographic method, which proved the reliability of the test strips. The limits of detection for the post-harvest and first-harvest tobacco samples were 0.1 mg/kg, and the test strips developed in this study are suitable for mass testing in tobacco laboratories with good application prospects because of their short detection time, simple pre-treatment and detection methods.

Development of Colloidal Gold Test Strips for the Detection of Triadimenol Residues in Tobacco.

The rapid and accurate determination of triadimenol residues is of great significance. In this study, based on the advantages of high efficiency, rapidity, reliability, simplicity and low cost of immunology, a test strip product for the rapid detection of triadimenol residues in tobacco was designed based on the optimization of conditions such as pH and dosage of diluent, concentration of antibody stock solution, type of confining solution and complex solution, with high specificity, accuracy and The results of 20 samples of fresh and first roasted tobacco were all consistent with the method of gas chromatography, which proved the reliability of the test strips. The detection limit for fresh and roasted tobacco was 5 mg/kg, and the test strips developed in this study are suitable for mass testing of tobacco samples in tobacco-related laboratories because of their short detection time, simple pre-treatment and detection methods, and good application prospects.

Rapid quantitative detection of chloramphenicol in three food products by lanthanide-labeled fluorescent-nanoparticle immunochromatographic strips.

A rapid and sensitive fluorescence-based immunochromatographic test (ICT) was successfully developed to determine chloramphenicol (CAP) levels in three food products. In this method, lanthanide fluorescent microspheres were used as a label to detect CAP in food samples within 30 min quantitatively, and the result was displayed on the test strip reader. After optimizing detection conditions, the detection limit (LOD) for the three food products was 0.048-0.073 ng g-1, and the half-maximal inhibitory concentration (IC50) was 0.27 ng mL-1. Six other veterinary drugs were detected using the test strip, and no cross-reactivity was observed, indicating that the specificity of the method was satisfactory. This method was also successfully applied to determine CAP in honey, egg, and fish samples, with recoveries ranging from 78.73% to 121.12%. The results demonstrated that this test strip had high sensitivity and specificity, and could be used for field detection within 30 min.

Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds.

Membrane-based lateral flow immunochromatographic strips (ICSs) are useful tools for low-cost self-diagnosis and have been efficiently applied to toxin, physiological index and clinical biomarker detection. In this protocol, we provide a detailed description of the steps to develop a rapid, sensitive and quantitative lateral-flow immunoassay (using AuNPs as a marker and mAbs as a probe). The procedure describes the preparation and characterization of colloidal gold, synthesis of the AuNP-mAb conjugate, assembly of the immunochromatographic strip, and methodological investigation of the assay. The results showed that the final strips can be further utilized for the rapid and convenient self-diagnosis of a small molecule, which may provide an alternative tool in the rapid and precise analysis of physiological and biological indices.

A simple pooling salivary test for SARS-CoV-2 diagnosis: A Columbus' egg?

Saliva is an appropriate specimen for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) diagnosis. The possibility of pooling samples of saliva, using non-invasive bibula strips for sampling, was explored employing Bovine coronavirus (BCoV) spiked saliva. In laboratory, up to 30 saliva-soaked strips were pooled in a single tube with 2 mL of medium. After quick adsorption with the medium and vortexing, the liquid was collected and tested with a quantitative molecular assay to quantify viral RNA genome copies. On testing of single and pooled strips, the difference between the median threshold cycles (Ct) value of test performed on the single positive saliva sample and the median Ct value obtained on the pool of 30 strips, was 3.21 cycles. Saliva pooling with bibula strips could allow monitoring of COVID-19 on a large scale, reducing costs for the health bodies in terms of medical material and skilled personnel. Finally, saliva sampling is noninvasive and less traumatic than nasopharyngeal swabs and can be self-collected.

One-step time-resolved fluorescence microsphere immunochromatographic test strip for quantitative and simultaneous detection of DON and ZEN.

Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that contaminate a wide range of grains and crops. In this study, a one-step time-resolved single-channel immunochromatographic test strip based on europium ion polystyrene fluorescence microspheres was first developed for sensitive and quantitative detection of DON and ZEN. The concentration of the artificial antigen and the mass ratio of the monoclonal antibody to fluorescent microspheres for conjugation were optimized to simplify the sample addition process during immunochromatographic assay and improve the on-site detection efficiency. The limits of detection (LOD) of the single-channel immunochromatographic test strip for DON and ZEN detection were 0.17 and 0.54 µg/L, respectively. Meanwhile, the dual-channel immunochromatographic test strip was designed to simultaneously detect DON and ZEN, with LODs of 0.24 and 0.69 µg/L achieved for DON and ZEN, respectively. The developed test strips also yielded recovery results consistent with that obtained by LC-MS/MS for DON and ZEN detection in real samples of wheat and corn flour, confirming the practicability and reliability of the test strip. The developed immunochromatographic test strips realize quick and sensitive detection of DON and ZEN, exhibiting potential for broad applications in the point-of-care testing platform of multiple mycotoxins in agricultural products. Graphic abstract.

Strip modification and alternative architectures for signal amplification in nanoparticle-based lateral flow assays.

Nanoparticle (NP)-based lateral flow assay (LFA) technology has outstanding characteristics that make it ideal for point-of-care bioanalytical applications. However, LFAs still have important limitations, especially related to sensitivity, which is in general worse than that of other well-established bioassays such as ELISA or PCR. Many efforts have been made for enhancing the sensitivity of LFAs, mainly actuating on the nanoparticle labels and on alternative optical detection modes. However, strip pads modification for such a purpose is an incipient vast field of research. This article gives a brief overview on the recent advances proposed for signal amplification actuating on different pads and the general architecture of the LFA strips. Such strategies offer universal tools that can be adapted to any LFA, independently of the kind of sample, analyte, and label. The principles of the different strategies developed to achieve novel signal amplification and sensitive detection are discussed, and some examples of relevant approaches are highlighted, together with future prospects and challenges.

Visual and quantitative detection of E. coli O157:H7 by coupling immunomagnetic separation and quantum dot-based paper strip.

Simple and visual quantitative detection of foodborne pathogens can effectively reduce the outbreaks of foodborne diseases. Herein, we developed a simple and sensitive quantum dot (QD)-based paper device for visual and quantitative detection of Escherichia coli (E. coli) O157:H7 based on immunomagnetic separation and nanoparticle dissolution-triggered signal amplification. In this study, E. coli O157:H7 was magnetically separated and labeled with silver nanoparticles (AgNPs), and the AgNP labels can be converted into millions of Ag ions, which subsequently quench the fluorescence of QDs in the paper strip, which along with the readout can be visualized and quantified by the change in length of fluorescent quenched band. Owing to the high capture efficiency and effective signal amplification, as low as 500 cfu mL-1 of E. coli O157:H7 could be easily detected by naked eyes. Furthermore, this novel platform was successfully applied to detect E. coli O157:H7 in spiked milk samples with good accuracy, indicating its potential in the detection of foodborne pathogens in real samples.

Implementing an integrated multi-technology platform for drug checking: Social, scientific, and technological considerations.

The illicit drug overdose crisis in North America continues to devastate communities with fentanyl detected in the majority of illicit drug overdose deaths. The COVID-19 pandemic has heightened concerns of even greater unpredictability in the drug supplies and unprecedented rates of overdoses. Portable drug-checking technologies are increasingly being integrated within overdose prevention strategies. These emerging responses are raising new questions about which technologies to pursue and what service models can respond to the current risks and contexts. In what has been referred to as the epicenter of the overdose crisis in Canada, a multi-technology platform for drug checking is being piloted in community settings using a suite of chemical analytical methods to provide real-time harm reduction. These include infrared absorption, Raman scattering, gas chromatography with mass spectrometry, and antibody-based test strips. In this Perspective, we illustrate some advantages and challenges of using multiple techniques for the analysis of the same sample, and provide an example of a data analysis and visualization platform that can unify the presentation of the results and enable deeper analysis of the results. We also highlight the implementation of a various service models that co-exist in a research setting, with particular emphasis on the way that drug checking technicians and harm reduction workers interact with service users. Finally, we provide a description of the challenges associated with data interpretation and the communication of results to a diverse audience.

What Is the Optimal Timing for Reading the Leukocyte Esterase Strip for the Diagnosis of Periprosthetic Joint Infection?

BACKGROUND: The leucocyte esterase (LE) strip test often is used to diagnose periprosthetic joint infection (PJI). In accordance with the manufacturer's directions, the LE strip test result is read 3 minutes after exposing it to joint fluid, but this has not been supported by robust research. Moreover, we have noted that the results of the LE strip test might change over time, and our previous studies have found that centrifugation causes the results of the LE strip test to degrade. Still, there is no evidence-based recommendation as to when to read the LE strip test to maximize diagnostic accuracy, in general, and the best reading times for the LE strip test before and after centrifugation need to be determined separately, in particular. QUESTIONS/PURPOSES: (1) What is the optimal timing for reading LE strip test results before centrifugation to diagnose PJI? (2) What is the optimal timing for reading LE strip test results after centrifugation to diagnose PJI? METHODS: This study was a prospective diagnostic trial. In all, 120 patients who were scheduled for revision arthroplasty and had signs of infection underwent joint aspiration in the outpatient operating room between July 2018 and July 2019 and were enrolled in this single-center study. For inclusion, patients must have had a diagnosis of PJI or nonPJI, valid synovial fluid samples, and must not have received antibiotics within 2 weeks before arthrocentesis. As such, 36 patients were excluded; 84 patients were included for analysis, and all 84 patients agreed to participate. The 2018 International Consensus Meeting Criteria (ICM 2018) was used for the classification of 49 patients with PJI (score ≥ 6) and 35 without PJI (score ≤ 2). The classification was used as the standard against which the different timings for reading LE strips were compared. All patients without PJI were followed for more than 1 year, during which they did not report the occurrence of PJI. All patients were graded against the diagnostic criteria regardless of their LE strip test results. In 83 patients, one drop of synovial fluid (50 µL) was applied to LE strips before and after centrifugation, and in one patient (without PJI), the sample was not centrifuged because the sample volume was less than 1.5 mL. The results of the strip test were read on an automated colorimeter. Starting from 1 minute after centrifugation, these strips were automatically read once every minute, 15 times (over a period of 16 minutes), and the results were independently recorded by two observers. Results were rated as negative, ±, 1+, and 2+ upon the machine reading. Grade 2+ (dark purple) was used as the threshold for a positive result. An investigator who was blinded to the study performed the statistics. Optimal timing for reading the LE strip before and after centrifugation was determined by using receiver operative characteristic (ROC) analysis. The specificity, sensitivity, and positive predictive and negative predictive values were calculated for key timepoints. RESULTS: Before centrifugation, the area under the curve was the highest when the results were read at 5 minutes (0.90 [95% CI 0.83 to 0.98]; sensitivity 0.88 [95% CI 0.75 to 0.95]; specificity 0.89 [95% CI 0.72 to 0.96]). After centrifugation, the area under the curve was the highest when the results were read at 10 minutes (0.92 [95% CI 0.86 to 0.98]; sensitivity 0.65 [95% CI 0.50 to 0.78]; specificity 0.97 [95% CI 0.83 to 1.00]). CONCLUSION: The LE strip test results are affected by time and centrifugation. For samples without centrifugation, we found that 5 minutes after application was the best time to read LE strips. We cannot deny the use of centrifuges because this is an effective way to solve the sample-mingling problem at present. We recommend 10 minutes postapplication as the most appropriate time to read LE strips after centrifugation. Multicenter and large-sample size studies are warranted to further verify our conclusion. LEVEL OF EVIDENCE: Level II, diagnostic study.

Desempenho de diferentes marcas de tiras reagentes relativo à densidade e à análise química da urina de cães e gatos / Performance of different brands of reagent strips for density and chemical analysis of dog and cat urine

Objetivou-se verificar a compatibilidade entre diferentes marcas de tiras reagentes para urinálise, tanto de uso veterinário, como de uso humano, e confrontar os parâmetros semiquantitativos desse instrumento com métodos quantitativos. Para isso, foram analisadas 77 amostras frescas de urina de cães e gatos e testados 04 modelos de tiras reagentes. Quanto à densidade urinária, houve correlação razoável entre os métodos quantitativo e semiquantitativo naquelas amostras com pH ácido, mas não naquelas com pH neutro ou alcalino. Quanto à concentração proteica, houve similaridade de 53,3% a 83,3% entre as marcas testadas e quando comparadas com a análise fotométrica houve uma correlação razoável (rs = 0,69752 a 0,75074). Em ponto de corte de 15mg/dL de proteína, a sensibilidade da tira reagente foi 82,5% e 100% para urina canina e felina, respectivamente. No tocante à hematúria, houve divergência razoável entre a sedimentoscopia e as diferentes marcas de tiras reativas. Quanto à piúria, há uma baixa sensibilidade das tiras em relação às amostras caninas com muitos resultados falso-negativos (33% a 75%), enquanto em amostras felinas a sensibilidade foi de 100%. Assim, independente da marca, as tiras reagentes devem servir apenas como teste rápido de triagem, sendo mais apropriado o uso de métodos quantitativos na avaliação clínica do paciente a partir da urinálise.

The aim was to verify the compatibility between different brands of urinary dipsticks, for both human and veterinary use, and to compare the semiquantitative parameters of this instrument with quantitative methods. For this, 77 fresh samples of urine from dogs and cats were analyzed e and 04 models of reagent strips were tested. Regarding urinary density, a reasonable correlation was observed between the quantitative and semiquantitative methods in those samples with acidic pH, which did not occur in those with neutral or alkaline pH. Regarding the protein concentration, there was similarity from 53.3% to 83.3% between the brands and in the comparative analysis between the control strip and the photometric analysis, there was a reasonable correlation (rs = 0.69752 to 0.75074). In cut-off point of 15mg/dL protein, the sensitivity of the reagent strip was 82.5% and 100% for canine and feline urine, respectively. Regarding hematuria, there was a reasonable divergence of results between sedimentation and tested dipsticks. As for pyuria, there is a low sensitivity of the strips in relation to canine samples with many false negative results (33% to 75%), while in feline samples the sensitivity was 100%. Thus, regardless of the brands, the reagent strips should serve only as a rapid screening test, while the use of quantitative methods in the clinical evaluation of the patient from urinalysis is more appropriate.

Controlled copper in situ growth-amplified lateral flow sensors for sensitive, reliable, and field-deployable infectious disease diagnostics.

A polyethyleneimine (PEI)-assisted copper in-situ growth (CISG) strategy was proposed as a controlled signal amplification strategy to enhance the sensitivity of gold nanoparticle-based lateral flow sensors (AuNP-LFS). The controlled signal amplification is achieved by introducing PEI as a structure-directing agent to regulate the thermodynamics of anisotropic Cu nanoshell growth on the AuNP surface, thus controlling shape and size of the resultant AuNP@Cu core-shell nanostructures and confining free reduction and self-nucleation of Cu2+ for improved reproducibility and decreased false positives. The PEI-CISG-enhanced AuNP-LFS showed ultrahigh sensitivities with the detection limits of 50 fg mL-1 for HIV-1 capsid p24 antigen and 6 CFU mL-1 for Escherichia coli O157:H7. We further demonstrated its clinical diagnostic efficacy by configuring PEI-CISG into a commercial AuNP-LFS detection kit for SARS-CoV-2 antibody detection. Altogether, this work provides a reliable signal amplification platform to dramatically enhance the sensitivity of AuNP-LFS for rapid and accurate diagnostics of various infectious diseases.

A point-of-care selenium nanoparticle-based test for the combined detection of anti-SARS-CoV-2 IgM and IgG in human serum and blood.

COVID-19 is a widespread and highly contagious disease in the human population. COVID-19 is caused by SARS-CoV-2 infection. There is still a great demand for point-of-care tests for detection, epidemic prevention and epidemiological investigation, both now and after the epidemic. We present a lateral flow immunoassay kit based on a selenium nanoparticle-modified SARS-CoV-2 nucleoprotein, which detects anti-SARS-CoV-2 IgM and anti-SARS-CoV-2 IgG in human serum, and the results can be read by the naked eye in 10 minutes. We expressed and purified the SARS-CoV-2 nucleoprotein in HEK293 cells, with a purity of 98.14% and a concentration of 5 mg mL-1. Selenium nanoparticles were synthesized by l-ascorbic acid reduction of seleninic acid at room temperature. After conjugation with the nucleoprotein, a lateral flow kit was successfully prepared. The IgM and IgG detection limits of the lateral flow kit reached 20 ng mL-1 and 5 ng mL-1, respectively, in human serum. A clinical study sample comprising 90 COVID-19-diagnosed patients and 263 non-infected controls was used to demonstrate a sensitivity and specificity of 93.33% and 97.34%, respectively, based on RT-PCR and clinical results. No cross-reactions with rheumatoid factor and positive serum for anti-nuclear antibodies, influenza A, and influenza B were observed. Moreover, the lateral flow kit remained stable after storage for 30 days at 37 °C. Our results demonstrate that the selenium nanoparticle lateral flow kit can conveniently, rapidly, and sensitively detect anti-SARS-CoV-2 IgM and IgG in human serum and blood; it can also be suitable for the epidemiological investigation of COVID-19.

Rapid detection of brucellosis using a quantum dot-based immunochromatographic test strip.

Novel diagnostic tools are a major challenge for brucellosis research, especially in developing countries. Herein, we established a handheld quantum dot (QD) immunochromatographic device for the fast detection of brucellosis antibodies in the field. Total bacterial protein extracted from Brucella 104M served as labelling and coating antigen. QD labelling and immunochromatography methods were used to optimise reaction conditions, labelling conditions, reaction temperature and storage temperature. QD test strips were employed to test brucellosis serum to determine their sensitivity, specificity and stability. Test strips were compared with Rose Bengal test, standard agglutination test and colloidal gold immunochromatographic assay. Labelled Brucella total protein displayed good specificity and no cross-reactivity. The concentration of labelled total bacterial protein was 3.9 mg/ml, the coating concentration was 2.0 mg/ ml, and the serum titre with the lowest detection sensitivity was 1:25. The optimal reaction temperature for the test strip was 25-30°C. The test strip was stable after storage at room temperature and the repeatability was high, with a coefficient of variation of 4.0%. After testing 199 serum samples, the sensitivity of the QD test strip was 98.53%, the specificity was 93.57%, and the coincidence rate with the standard agglutination test was 96.98%. The developed QD immunochromatographic method can be used for rapid detection and preliminary screening of brucellosis in the field.

Contamination of Zearalenone from China in 2019 by a Visual and Digitized Immunochromatographic Assay.

Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of detection (LOD) of 0.50 ng/mL with visual judgment and could be completely inhibited within 5 min at 3.0 ng/mL ZEN. The quantitative detection model had a lower LOD of 0.25 ng/mL, and ZEN could be accurately and digitally detected from 0.25-4.0 ng/mL. The ICA method had a high sensitivity, specificity, and accuracy for on-site ZEN detection. For investigation of the authentic samples, the ZEN-positive rate was 62.6%, and the ZEN-positive levels ranged from 2.7 to 867.0 ng/g, with an average ZEN-positive level being 85.0 ng/g. Of the ZEN-positive samples, 6.0% exceeded the values of the limit levels. The ZEN-positive samples were confirmed to be highly correlated using LC-MS/MS (R2 = 0.9794). This study could provide an efficiency and accuracy approach for ZEN in order to achieve visual and digitized on-site investigation. This significant information about the ZEN contamination levels might contribute to monitoring mycotoxin occurrence and for ensuring food safety.

Diagnostic methods and potential portable biosensors for coronavirus disease 2019.

Timely detection and diagnosis are urgently needed to guide epidemiological measures, infection control, antiviral treatment, and vaccine research. In this review, biomarkers/indicators for diagnosis of coronavirus disease 2019 (COVID-19) or detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the environment are summarized and discussed. It is concluded that the detection methods targeting antibodies are not suitable for screening of early and asymptomatic cases since most patients had an antibody response at about 10 days after onset of symptoms. However, antibody detection methods can be combined with quantitative real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) to significantly improve the sensitivity and specificity of diagnosis, and boost vaccine research. Fast, sensitive and accurate detection methods targeting antigens need to be developed urgently. Various specimens for diagnosis or detection are compared and analyzed. Among them, deep throat saliva and induced sputum are desired for RT-qPCR test or other early detection technologies. Chest computerized tomography (CT) scan, RT-qPCR, lateral flow immunochromatographic strip (LFICS) for diagnosis of COVID-19 are summarized and compared. Specially, potential electrochemical (EC) biosensor, surface enhanced Raman scattering (SERS)-based biosensor, field-effect transistor (FET)-based biosensor, surface plasmon resonance (SPR)-based biosensor and artificial intelligence (AI) assisted diagnosis of COVID-19 are emphasized. Finally, some commercialized portable detection device, current challenges and future directions are discussed.

Terminal-conjugated non-aggregated constraints of gold nanoparticles on lateral flow strips for mobile phone readouts of enrofloxacin.

Antibiotics abuse now poses a global threat to public health. Monitoring their residual levels as well as metabolites are of great importance, still challenges remain in in situ tracing during the circulation. Herein, taking the typical antibacterial Enrofloxacin (ENR) as a subject, a paper-based aptasensor was tailored by manipulating a duo of aptameric moieties to "sandwich" the target in a lateral-flow regime. To visualize the tight-binding sandwich motif more vividly, an irregular yet robust DNA-bridged gold nanoparticles (AuNPs) proximity strategy was developed with recourse to terminal deoxynucleotidyl transferase, of which the nonaggregate constraining feature was unveiled via optical absorption and scanning probe topography. This complex performed exceptionally better in the test strip context than single-particle tags, leading to an enhanced on-chip focusing. Rather than qualitative color developing, further efforts were taken to visualize the readouts in a quantitative manner by exploiting the smartphone camera for pattern recognition along with data processing in a professional App. Overall, this prototyped contraption realized a rapid and ultrasensitive quantification of ENR down to 0.1 µg/L along with a broad linear range over 5 orders of magnitude, plus excellent selectivity and precision even for real samples. Such innovative fusion across DNA-structured nanomanufacturing and intelligent perception provides a prospective and invigorating solution to point-of-care inspection.

Inkjet-printed electrochemically reduced graphene oxide microelectrode as a platform for HT-2 mycotoxin immunoenzymatic biosensing.

The design and application of an inkjet-printed electrochemically reduced graphene oxide microelectrode for HT-2 mycotoxin immunoenzymatic biosensing is reported. A water-based graphene oxide ink was first formulated and single-drop line working microelectrodes were inkjet-printed onto poly(ethylene 2,6-naphthalate) substrates, with dimensions of 78 µm in width and 30 nm in height after solvent evaporation. The printed graphene oxide microelectrodes were electrochemically reduced and characterized by Raman and X-ray photoelectron spectroscopies in addition to microscopies. Through optimization of the electrochemical reduction parameters, differential pulse voltammetry were performed to examine the sensing of 1-naphthol (1-N), where it was revealed that reduction times had significant effects on electrode performance. The developed microelectrodes were then used as an immunoenzymatic biosensor for the detection of HT-2 mycotoxin based on carbodiimide linking of the microelectrode surface and HT-2 toxin antigen binding fragment of antibody (anti-HT2 (10) Fab). The HT-2 toxin and anti-HT2 (10) Fab reaction was reported by anti-HT2 immune complex single-chain variable fragment of antibody fused with alkaline phosphatase (anti-IC-HT2 scFv-ALP) which is able to produce an electroactive reporter - 1-N. The biosensor showed detection limit of 1.6 ng ∙ mL-1 and a linear dynamic range of 6.3 - 100.0 ng ∙ mL-1 within a 5 min incubation with 1-naphthyl phosphate (1-NP) substrate.