Three-dimensional assembly and disassembly of Fe3O4-decorated porous carbon nanocomposite with enhanced transversal relaxation for magnetic resonance sensing of bisphenol A.

The design and construction of a novel magnetic resonance sensor (MRS) is presented for bisphenol A (BPA) detection. The MRS has been built based on the core component of magnetic Fe3O4 nanoparticles (~ 40 nm), which were uniformly distributed in nanoporous carbon (abbreviated as Fe3O4@NPC). The synthesis was derived from the calcination of the metal organic framework (MOF) precursor of Fe-MIL-101 at high temperature. Fe3O4@NPC was confirmed with enhanced transversal relaxation with r2 value of 118.2 mM-1 s-1, which was around 1.7 times higher than that of the naked Fe3O4 nanoparticle. This enhancement is attributed to the excellent proton transverse relaxation rate of Fe3O4@NPC caused by the reduced self-diffusion coefficient of water molecules in the vicinity of Fe3O4 nanoparticles in the nanoporous carbon. BPA antibody (Ab) and antigen (Ag)-ovalbumin (OVA) were immobilized onto the Fe3O4@NPC to form Ab-Fe3O4@NPC and Ag-Fe3O4@NPC, respectively. These two composites can cause the three-dimensional assembly of Fe3O4@NPC via immunological recognition. The presence of BPA can compete with antigen-OVA to combine with Ab-Fe3O4@NPC, thereby breaking the assembly process (disassembly). The difference in the change of the T2 value before and after adding BPA can thus be used to monitor BPA. The proposed MRS not only revealed a wide linear range of BPA concentration from 0.05 to 50 ng mL-1 with an extremely low detection limit of 0.012 ng mL-1 (S/N = 3), but also displayed high selectivity towards matrix interferences. The recoveries of BPA ranged from 95.6 to 108.4% for spiked tea π, and 93.4 to 104.7% for spiked canned oranges samples, respectively, and the RSD (n = 3) was less than 4.4% for 3 successive assays. The versatility of Fe3O4@NPC with customized relaxation responses provides the possibility for the adaptation of magnetic resonance platforms for food safety development. The magnetic Fe3O4 nanoparticles are uniformly dispersed in the nanoporous carbon (Fe3O4@NPC), which derived from the calcinating of the metal organic framework (MOF) precursor of Fe-MIL-101. And the magnetic Fe3O4@NPCs are adopted for the construction of magnetic resonance sensor (MRS) for bisphenol A (BPA) detection.

Molybdenum disulfide-gold nanoparticle nanocomposite in field-effect transistor back-gate for enhanced C-reactive protein detection.

Nanofabricated gold nanoparticles (Au-NPs) on MoS2 nanosheets (Au-NPs/MoS2) in back-gated field-effect transistor (BG-FET) are presented, which acts as an efficient semiconductor device for detecting a low concentration of C-reactive protein (C-RP). The decorated nanomaterials lead to an enhanced electron conduction layer on a 100-µm-sized transducing channel. The sensing surface was characterized by Raman spectroscopy, ultraviolet-visible spectroscopy (UV-Vis), atomic force microscopy (AFM), scanning electron microscopy (SEM), and high-power microscopy (HPM). The BG-FET device exhibits an excellent limit of detection of 8.38 fg/mL and a sensitivity of 176 nA/g·mL-1. The current study with Au-NPs/MoS2 BG-FET displays a new potential biosensing technology; especially for integration into complementary metal oxide (CMOS) technology for hand-held future device application.

Semiquantitative immunochromatographic colorimetric biosensor for the detection of dexamethasone based on up-conversion fluorescent nanoparticles.

A low-cost bifunctional immunochromatographic colorimetric biosensor was developed that can be read visually or by using an optical density scanner. Five test lines (T lines) coated with different antigens were set on a nitrocellulose (NC) membrane to indicate the concentration of analyte. This method was applied for the detection of dexamethasone. The corresponding detection range was 0.1-9 ng mL-1, and the detection limit for dexamethasone in food supplements and cosmetic samples was 2.0 µg kg-1. For visual inspection of the colour the quantitative relative error range between the proposed method and liquid chromatography was -62 to -25%, with a detection time of only 10 min. More accurate assay results were obtained by using an optical density scanner with the relative error range of -31 to 20%. The results indicated that the proposed method has the potential of application for rapid and efficient screening of dexamethasone in cosmetics and food supplements. Graphical abstract.

Gold nanoparticle-modified black phosphorus nanosheets with improved stability for detection of circulating tumor cells.

Gold nanoparticle (AuNP)-anchored BP nanosheets were synthesized through in situ growth of AuNPs onto BP. Due to the strong chelating ability of P or phosphorus oxides with AuNPs, the stability of BP is improved. As proof-of-concept demonstration of the functionalized BP, electrochemical detection of circulating tumor cells (CTCs) based on BP@AuNPs@aptamer as a probe combined with immunomagnetic separation is reported. The aptamer can specifically bind with CTCs, while the phosphorus oxides including phosphite ion and phosphate ion (PxOy species) on BP and aptamer can react with molybdate to generate an electrochemical current, leading to dual signal amplification. The biosensor is applied to MCF-7 cell detection and displays good analytical performance with a detection limit of 2 cell mL-1. Furthermore, the practicality of this biosensor was validated through sensitive determination of MCF-7 cells in human blood. Therefore, the reported biosensor could be applied to detect other biomarkers, offering an ultrasensitive strategy for clinical diagnostics. Graphical abstract Electrochemical detection of circulating tumor cells based on gold nanoparticle-modified black phosphorus nanosheets is reported.

Rapid screening and quantitative detection of Salmonella using a quantum dot nanobead-based biosensor.

The continuing hurdle of developing foodborne pathogen detection techniques is that compromises must be made among simplicity, portability, speed, sensitivity, and quantitation. Herein, we fabricated quantum dot nanobeads (QDNS) by a layer-by-layer assembly of quantum dots on the surface of polymer nanospheres. QDNS exhibited higher fluorescence intensity than the quantum dots at the same particle number. Based on the quantum dot nanobeads as the signal reporter, a quantitative lateral flow immunoassay was demonstrated for Salmonella typhimurium detection with improved sensitivity, specificity and accuracy. A visual detection limit of 5 × 103 CFU mL-1Salmonella typhimurium within 10 min has been proved and demonstrated. Additionally, higher concentrations of non-Salmonella typhimurium bacteria have negligible effects on the detection of Salmonella typhimurium. The results of 50 single blind tests by 10 testers suggested that the assay exhibited 100% accuracy. The results illustrate that the assay provides a balance among simplicity, speed, sensitivity and accuracy, and it can be a favorable alternative for Salmonella typhimurium screening in various samples.

Targeted removal of leukemia cells from the circulating system by whole-body magnetic hyperthermia in mice.

Until now, magnetic hyperthermia was used to remove solid tumors by targeting magnetic nanoparticles (MNPs) to tumor sites. In this study, leukemia cells in the bloodstream were directly removed by whole-body hyperthermia, using leukemia cell-specific MNPs. An epithelial cellular adhesion molecule (EpCAM) antibody was immobilized on the surface of MNPs (EpCAM-MNPs) to introduce the specificity of MNPs to leukemia cells. The viability of THP1 cells (human monocytic leukemia cells) was decreased to 40.8% of that in control samples by hyperthermia using EpCAM-MNPs. In AKR mice, an animal model of lymphoblastic leukemia, the number of leukemia cells was measured following the intravenous injection of EpCAM-MNPs and subsequent whole-body hyperthermia treatment. The result showed that the leukemia cell number was also decreased to 43.8% of that without the treatment of hyperthermia, determined by Leishman staining of leukemia cells. To support the results, simulation analysis of heat transfer from MNPs to leukemia cells was performed using COMSOL Multiphysics simulation software. The surface temperature of leukemia cells adhered to EpCAM-MNPs was predicted to be increased to 82 °C, whereas the temperature of free cells without adhered MNPs was predicted to be 38 °C. Taken together, leukemia cells were selectively removed by magnetic hyperthermia from the bloodstream, because EpCAM-modified magnetic particles were specifically attached to leukemia cell surfaces. This approach has the potential to remove metastatic cancer cells, and pathogenic bacteria and viruses floating in the bloodstream.

Cooperation Mode of Outer Surface and Inner Space of Nanochannel: Separation-Detection System Based on Integrated Nanochannel Electrode for Rapid and Facile Detection of Salmonella.

Nanochannels hold great prospects in intelligent systems; however, current research focuses on the inner space of the nanochannel while the outer surface is rarely explored. Here, we report on a cooperation mode of the outer surface and inner space of the nanochannel using an integrated nanochannel-electrode (INCE) and its application as a separation-detection system for rapid and facile detection of foodborne bacteria. Unlike conventional nanochannel systems, the INCE integrates two electrodes as a sensitive electrochemical interface and the nanochannel itself as nanofilter, generating a novel separation-detection system. The system is examined in a biosensing strategy based on magnetic nanoparticles (MNPs). Salmonella typhimurium (St) is taken as the target due to its severe threat to human health and food safety. By electrochemically probing the MNPs-St complex themselves on the surface of INCE, this method eliminates the requirement on additional signal labels. The biosensor presents a linear detection range from 102 to 107 CFU mL-1 and a limit of detection of 50 CFU mL-1, being comparable or even better than those of analogues with complicated signal amplification designs. Moreover, the biosensor exhibits good specificity against four types of interfering bacteria. This concept may bring new insight into the development of nanochannel research and contribute a new way to the fields of separation and detection.

Dual-signal sandwich electrochemical immunosensor for amyloid ß-protein detection based on Cu-Al2O3-g-C3N4-Pd and UiO-66@PANI-MB.

Combining of amperometric and square wave voltammetric methods (SWV), the dual-signal sandwich electrochemical immunosensor was developed for quantitative determination of amyloid ß-protein (Aß). Cu was doped into Al2O3 lattice (Cu-Al2O3) and reacts with graphite carbon nitride (g-C3N4) to generate Cu-Al2O3-g-C3N4 with internal dual-reaction center structure, which has good catalytic properties of hydrogen peroxide (H2O2). Subsequently, palladium nanoparticles (Pd NPs) was introduced into Cu-Al2O3-g-C3N4 (Cu-Al2O3-g-C3N4-Pd) that not only synergistically catalyzed H2O2 but also immobilized anti-Aß (Ab1) via Pd-NH2. The Cu-Al2O3-g-C3N4-Pd was used as matrix material to modify the electrode, which can produce obviously electrochemical signals through Amperometry i-t curve. Meanwhile, the Zr6O4(OH)4(CO2)12 (UiO-66) modified with polyaniline (PANI) has the large specific surface, good conductivity and adsorption capacity, which can support methylene blue (MB) as signal label of anti-Aß (Ab2). Therefore, the UiO-66@PANI-MB can provide an obviously electrochemical signal about MB through SWV. Under optimal conditions, the dual-signal sandwich electrochemical immunosensor has salient analytical performance and both signal platforms provide more accurate results. The linear range of detection obtained by the immunosensor was 10 fg/mL-100 ng/mL, and the detection limit was 3.3 fg/mL. This method not only provided a reliable guarantee for the experimental detection but also provided an effective strategy for the detection of other biological.

Low fouling and ultrasensitive electrochemical immunosensors with dual assay methods based on Fe3O4 magnetic nanoparticles.

Low fouling electrochemical immunosensors with both "signal-off" and "signal-on" analytical methods were developed for the highly sensitive and efficient detection of cancer antigen 15-3 (CA 15-3) in human serum samples. The antifouling sensing interfaces were constructed by assembling multifunctional polyethylene glycol on gold electrodes, followed by covalent conjugation with CA 15-3 antibody. Pure antigens and Fe3O4@Ag will competitively bind to the immobilized antibody on the electrode. Fe3O4 magnetic nanoparticles attached to the working electrode and collected by a magnetic electrode were treated via electrochemical conversion to generate electroactive Prussian blue as a signal readout. Therefore, these two signals measured independently were complementary, and this design allowed one to choose the assay method according to real situations so as to ensure accuracy of the immunosensor. Moreover, owing to its good antifouling property, the immunosensor was capable of detecting CA 15-3 even in complex human serum samples, demonstrating potential application in quantitative analysis of real patient serum samples.

Magnetic particles-based enzyme immunoassay for rapid determination of secoiridoid glycoside, amarogentin.

Amarogentin (AG) is one of the bitter secoiridoid glycosides, which exerts various pharmacological activities as a bitter stomachic. Recently, there is an increasing demand for AG-containing plants in Japan due to their use as folk medicines and food additives; hence, it is crucial to develop analytical techniques that are specific for AG. In this study, a new magnetic particles-based enzyme immunoassay (MPs-EIA) using a specific monoclonal antibody against AG (MAb 1E9) for the rapid determination of AG in plants of the family Gentianaceae was described. AG directly immobilized onto magnetic particles (MPs) was used as a competitor for free AG against MAb 1E9, thereby increasing the surface area of the solid phase and decreasing the immunoreaction time. In addition, the blocking step required in case of the conventional enzyme-linked immunosorbent assay could be avoided in the proposed MPs-EIA, which enables an even more rapid performance for the immunoassay. In the developed MPs-EIA, AG exhibited linearity in the range of 15.6-500 ng mL-1, with a limit of detection of 8.58 ng mL-1. Validation analysis revealed that MPs-EIA is a sufficiently sensitive and rapid for the quantitative analysis of AG in plant samples. To the best of our knowledge, this is the first MPs-EIA that has been applied to plant samples.

Plasmonic Cu2- xS ySe1- y Nanoparticles Catalyzed Click Chemistry Reaction for SERS Immunoassay of Cancer Biomarker.

Nature enzyme-based immunoassays have been widely used in fundamental scientific research and clinical diagnosis. However, the limitations of natural enzyme, such as the low physical/chemical stability or susceptibility to protein denaturation, greatly restrained its applications. In this article, we reported a new enzyme-free SERS immunoassay by utilizing plasmonic Cu2- xS ySe1- y nanoparticles (NPs) as nanocatalyst to catalyze the click chemistry between the azido and alkynyl substrate which is used as the SERS signal reporter. The unique vibration of C≡C of alkynyl in the Raman-silent region (1800-2800 cm-1) is not overlapped with the signals of the other conventional Raman reporters or endogenous biological species, and thus it can make sure the enzyme-free SERS immunoassay has high selectivity and sensitivity. As a proof of concept, prostate-specific antigen (PSA), a biomarker of prostate cancer in blood, has been detected. The SERS immunoassay shows good analytical performance for PSA in the range of 3-120 ng mL-1, and it has been successfully applied to detect PSA in the serum samples of prostate cancer patients, proving that the proposed enzyme-free SERS immunoassay has great potential in the clinical diagnosis of cancer.

Fluorescence-based immunosensor using three-dimensional CNT network structure for sensitive and reproducible detection of oral squamous cell carcinoma biomarker.

A hierarchical three-dimensional network of carbon nanotubes on Si pillar substrate (3DN-CNTs) was developed for the accurate detection of oral squamous cell carcinoma (OSCC) in clinical saliva samples. The 3DN-CNTs were uniformly coated with a layer of aluminum oxides to enhance structural stability during biomarker detection. Cytokeratin-19 antigen (Cyfra 21-1) was utilized as a model biomarker of OSCC for fluorescence-based immunosensor using 3DN-CNTs (3DN-CNTs sensor). The 3DN-CNTs sensor enhances the sensitivity of Cyfra 21-1 detection by increasing the density of immobilized antibody through high surface area of 3DN-CNTs and enhancing the accessibility of biomolecules through the ordered pathway of hierarchical structure. The reliable detection limit for sensing of Cyfra 21-1 was estimated as in the level of 0.5 ng/mL and the quantitative estimation of Cyfra 21-1 was analyzed by 4-parameter logistic (4-PL) model for curve-fitting analysis. Clinical applicability of 3DN-CNTs sensor was evaluated through correlation with the commercially available electrochemiluminescence (ECL) detection system in the hospital. The assay results of the two systems for clinical saliva samples showed a good linear correlation. The 3DN-CNTs sensor offers great potential for accurate diagnosis of OSCC using Cyfra 21-1 biomarker in clinical fluids.

Immunoaffinity chromatography combined with tandem mass spectrometry: A new tool for the selective capture and analysis of brassinosteroid plant hormones.

Brassinosteroids (BRs) are plant-specific steroid hormones that play essential roles in the regulation of many important physiological processes in plant life. Their extremely low concentrations (~pmoles/g FW) in plant tissue and huge differences in polarity of individual members within the BR family hamper their detection and quantification. To address this problem, an immunoaffinity sorbent with broad specificity and high capacity for different BR metabolites containing a monoclonal antibody (mAb) against a BR spacer (20S)-2α,3α-dihydroxy-7-oxa-7α-homo-5α-pregnane-6-one-20 carboxylic acid (BR4812) was used for the rapid and highly selective isolation of endogenous BRs containing a 2α,3α-diol in ring A from minute plant samples. This enrichment procedure was successfully applied as a sample preparation method prior to quantitative analysis of BRs in real plant tissues by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Use of immunoaffinity chromatography (IAC) increased the sensitivity of the UHPLC-MS/MS analysis owing to improvements in the BR signal-to-noise ratio (S/N) and matrix factor (MF). Although MF values of BRs analyzed in classical samples ranged from 8.9% to 47.4%, MF values for the IAC purified samples reached 44.5-96.6%. Thus, the developed IAC-UHPLC-MS/MS approach was shown to be a simple, robust, effective and extremely fast procedure requiring minute amounts of plant samples suitable for the quantitative profiling of many BR metabolites, helping to overcome the major problems associated with their determination in very complex plant matrices.

High-sensitivity immunochromatographic assay for fumonisin B1 based on indirect antibody labeling.

OBJECTIVE: To develop a high-sensitivity immunochromatographic test for fumonisin B1 in plant extracts. RESULTS: Unlike conventional immunochromatographic tests, this assay is performed in two stages: competitive reaction with free specific antibodies and identifying immune complexes by their interaction with the anti-species antibody-conjugated gold nanoparticles. The use of a new geometry for the test strip membranes and a novel reagent application method ensures the proper order of these stages without additional manipulations. The contact of the ready-to-use test strip with the liquid sample suffices in initiating all stages of the assay and obtaining test results. The developed test was used on corn extracts; its instrumental limit of fumonisin B1 detection was 0.6 ng ml-1 at 15 min of assay duration. CONCLUSIONS: The proposed approach is flexible and can be used for a wide range of low molecular compounds. The use of anti-species antibody-conjugated gold nanoparticles in immunochromatography significantly facilitates the development of test systems by eliminating the need to synthesize and characterize the conjugates with specific antibodies for each new compound to be detected.

Field-effect transistor biosensors with two-dimensional black phosphorus nanosheets.

A black phosphorous (BP)-based field-effect transistor (FET) biosensor was fabricated by using few-layer BP nanosheets labeled with gold nanoparticle-antibody conjugates. BP nanosheets were mechanically exfoliated and used as the sensing/conducting channel in the FET, with an Al2O3 thin film as the dielectric layer for surface passivation. Antibody probes were conjugated with gold nanoparticles that were sputtered on the BP through surface functionalization. The sensor response was measured by the change in the BP's electrical resistance after antigens were introduced. The adsorbed antigens through specific antigen-antibody binding interactions induced a gate potential, thereby changing the drain-source current. The as-produced BP biosensor showed both high sensitivity (lower limit of detection ~10ng/ml) and selectivity towards human immunoglobulin G. Results from this study demonstrate the outstanding performance of BP as a sensing channel for FET biosensor applications.

Immunochromatographic determination of bacopaside I in biological samples.

We describe a novel immunochromatographic method for qualitative and quantitative analyses of bacopaside I, a bioactive constituent in Bacopa monnieri (L.) Wettst in biological samples. The assay was performed on polyethersulfone membrane using a polyclonal antibody raised against bacopaside I. The finalised method could quantitatively determine bacopaside I in the range of 31.3-1000.0ng and the detection and quantification limits were 1.0 and 31.3ng, respectively. The percentage recoveries of bacopaside I in blood and urine were nearly 100% indicating the accuracy of the extraction. The method was then applied for the determination of this compound in rat serum, urine and feces after an oral dose of 15mg/kg body weight. About 4% of the ingested dose of bacopaside I was detected in rat feces but none was detected in serum and urine which accorded with results from liquid chromatography tandem mass spectrometry. The accuracy, selectivity, sensitivity of the method are appropriate for in vivo pharmacokinetic studies.

Selective tools for the solid-phase extraction of Ochratoxin A from various complex samples: immunosorbents, oligosorbents, and molecularly imprinted polymers.

The evolution of instrumentation in terms of separation and detection has allowed a real improvement of the sensitivity and the analysis time. However, the analysis of ultra-traces of toxins such as ochratoxin A (OTA) from complex samples (foodstuffs, biological fluids…) still requires a step of purification and of preconcentration before chromatographic determination. In this context, extraction sorbents leading to a molecular recognition mechanism appear as powerful tools for the selective extraction of OTA and of its structural analogs in order to obtain more reliable and sensitive quantitative analyses of these compounds in complex media. Indeed, immunosorbents and oligosorbents that are based on the use of immobilized antibodies and of aptamers, respectively, and that are specific to OTA allow its selective clean-up from complex samples with high enrichment factors. Similar molecular recognition mechanisms can also be obtained by developing molecularly imprinted polymers, the synthesis of which leads to the formation of cavities that are specific to OTA, thus mimicking the recognition site of the biomolecules. Therefore, the principle, the advantages, the limits of these different types of extraction tools, and their complementary behaviors will be presented. The introduction of these selective tools in miniaturized devices will also be discussed.

Spectrum-Based Electrochemiluminescent Immunoassay with Ternary CdZnSe Nanocrystals as Labels.

Conventional electrochemiluminescence (ECL) research has been performed by detecting the total photons (i.e., the ECL intensity). Herein, systematic spectral exploration on the ECL of dual-stabilizers-capped ternary CdZnSe nanocrystals (NCs) and its sensing application were carried out on a homemade ECL spectral acquiring system. The ternary CdZnSe NCs could be repeatedly injected with electrons via some electrochemical ways and then result in strong cathodic ECL with the coupling of ammonium persulfate. ECL spectrum of the CdZnSe NCs was almost identical to corresponding photoluminescence spectrum, indicating that the excited states of CdZnSe NCs in ECL were essentially the same as those in photoluminescence. Importantly, after being labeled to the probe antibody (Ab2) of α-fetal protein (AFP) antigen, the ternary NCs in the Ab2|NCs conjugates could preserve their ECL spectrum very well. A spectrum-based ECL immunoassay was consequently proposed with the CdZnSe NCs as ECL tags and AFP as target molecules. The limit of detection is 0.010 pg/mL, with a signal-to-noise (S/N) ratio of 3, indicating a sensitive ECL sensing strategy that was different from the conventional ones. This work might open a pathway to the spectrally resolved ECL analysis with even-higher S/N ratios than the fluorescent analysis.

Quantifying Aflatoxin B1 in peanut oil using fabricating fluorescence probes based on upconversion nanoparticles.

Rare earth doped upconversion nanoparticles convert near-infrared excitation light into visible emission light. Compared to organic fluorophores and semiconducting nanoparticles, upconversion nanoparticles (UCNPs) offer high photochemical stability, sharp emission bandwidths, and large anti-Stokes shifts. Along with the significant light penetration depth and the absence of autofluorescence in biological samples under infrared excitation, these UCNPs have attracted more and more attention on toxin detection and biological labelling. Herein, the fluorescence probe based on UCNPs was developed for quantifying Aflatoxin B1 (AFB1) in peanut oil. Based on a specific immunity format, the detection limit for AFB1 under optimal conditions was obtained as low as 0.2ng·ml(-1), and in the effective detection range 0.2 to 100ng·ml(-1), good relationship between fluorescence intensity and AFB1 concentration was achieved under the linear ratios up to 0.90. Moreover, to check the feasibility of these probes on AFB1 measurements in peanut oil, recovery tests have been carried out. A good accuracy rating (93.8%) was obtained in this study. Results showed that the nanoparticles can be successfully applied for sensing AFB1 in peanut oil.

Multiarray on a test strip (MATS): rapid multiplex immunodetection of priority potato pathogens.

Multiarray on a test strip (MATS) was developed for the detection of eight important potato pathogens. The proposed assay combines the rapidity of immunochromatography with the high throughput of array techniques. The test zone of the immunochromatographic strip comprises ordered rows of spots containing antibodies specific for different potato pathogens. The assay benefits from the simplicity of immunochromatography; colored immune complexes form at the corresponding spots within the test zone. The presence and intensity of the coloration are used for identification of the target pathogens. The MATS was applied to the simultaneous detection of eight priority potato pathogens, characterized by the following limits of detection: 1 ng/mL for potato virus X and the ordinary type of potato virus Y, 10 ng/mL for potato virus M, 20 ng/mL for potato leaf roll virus, 40 ng/mL for necrotic-type potato virus Y, 100 ng/mL for potato virus S, 300 ng/mL for potato virus A, and 10(4) cells/mL for Clavibacter michiganensis subsp. sepedonicus. Analysis time was 15 min. The observed sensitivity of the MATS was comparable to the traditional enzyme-linked immunosorbent assay. The developed technique was tested on potato leaf extracts, and its efficiency for on-site control of the pathogens was confirmed in 100 % by commercial LFIA test strips. Graphical abstract Location of binding zones in the developed multiarray on a test strip (MATS) for simultaneous detection of eight pathogens.