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The Zika virus (ZIKV) is a great concern for global health due to its high transmission, including disseminating through blood, saliva, urine, semen and vertical transmission. In some cases, ZIKV has been associated with microcephaly, neurological disorders, and Guillain−Barré syndrome. There is no vaccine, and controlling the disease is a challenge, especially with the co-circulation of the Dengue virus, which causes a severe cross-reaction due to the similarity between the two arboviruses. Considering that electrochemical immunosensors are well-established, sensitive, and practical tools for diagnosis, in this study we developed a sensor platform with intrinsic redox activity that facilitates measurement readouts. Prussian blue (PB) has a great ability to form electrocatalytic surfaces, dispensing redox probe solutions in voltammetric measurements. Herein, PB was incorporated into a chitosan−carbon nanotube hybrid, forming a nanocomposite that was drop-casted on a screen-printed electrode (SPE). The immunosensor detected the envelope protein of ZIKV in a linear range of 0.25 to 1.75 µg/mL (n = 8, p < 0.01), with a 0.20 µg/mL limit of detection. The developed immunosensor represents a new method for electrochemical measurements without additional redox probe solutions, and it is feasible for application in point-of-care diagnosis., Competing Interests: The authors declare no conflict of interest.

An electrochemical immunosensor based on a nanohybrid film of carboxylated polypyrrole and amine nanoclay was developed for label-free detection of the human cardiac troponin T (cTnT). The nanohybrid film was formed in situ on the surface of the glassy carbon electrode, followed by the covalent immobilization of anti-troponin T antibodies by glutaraldehyde. Morphological and chemical characterizations of the nanohybrid film were performed by scanning electron microscopy and Fourier-transform infrared spectroscopy. Under the optimized conditions, a calibration curve for cTnT in spiked serum was obtained by square wave voltammetry, and a low limit of detection and quantification was achieved (0.35 and 1.05 pg mL -1 , respectively). This was the first time that this type of nanohybrid film was used in the development of an immunosensor for cTnT that proved to be a simple and efficient strategy for the manufacture of a label-free electrochemical device that could be applied in the diagnosis of acute myocardial infarction.

An ultrasensitive label-free electrochemical immunosensor was developed for hepatitis C antibodies (anti-HCV). Worldwide, it is estimated 71 million people have HCV infection in a chronic stage that may lead to cirrose and cancer. To achieve HCV elimination, health programs should include screening testing based on anti-HCV detection allowing the early-stage treatment. The immunosensor was based on a graphene oxide-polypyrrole (PPy-GO) film one-step electropolymerized on the electrode surface. Ultrasensitive anti-HCV detection was ensured by HCV antigen conjugated to biotin that was immobilized in a great amount on streptavidin-coated nanostructured surface. Analytical responses were obtained by anodic peaks from the square wave voltammetry in the presence of ferrocyanide/ferricyanide as a redox probe. This immunosensor exhibited a linear range from 2 to 14 ng mL -1 of anti-HCV and a limit of detection in the clinical range (1.63 ng mL -1 ). Furthermore, the immunosensor presented an efficient performance for the determination of anti-HCV in spiked serum samples, becoming this developed nanosensor as potential tools for early HCV diagnosis and screening., Supplementary Information: The online version contains supplementary material available at 10.1007/s10853-022-06992-5., Competing Interests: Conflict of interestAll authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022.)

Zika virus (ZIKV) infections were associated with neurological disorders only after the Brazilian outbreak in 2015. The lack of vaccines and precise diagnosis requires a precise method to detect ZIKV infection. This study aimed to evaluate three ZIKV recombinant proteins for the development of ZIKV infections. Here, it was purified stable recombinant ZIKV Capsid (r-ZIKV-c), non-structural proteins NS1 (r-ZIKV-NS1), and NS3 (r-ZIKV-NS3) for detection of the infection by ZIKV in blood sera of patients. A commercial polyclonal antibody recognized the r-ZIKV-NS1. Here, among three proteins, NS1 showed the best result for diagnostic purposes using serum samples, despite the high similarity with NS1 from DENV, and could differentiate the infections. The recombinant NS1 was used to produce a monoclonal antibody to differentiate between DENV and ZIKV NS1. As for recombinant proteins, the result for r-ZIKV-NS1 values showed 77% and 100% sensitivity and specificity, respectively, in the IgM assay. Our data showed the protein could successfully differentiate between sera of ZIKV infected patients from sera of those not infected with the virus and differentiate from sera of DENV infected patients. Thus, the generated recombinant proteins have great potential for serological diagnosis of ZIKV in Brazil, where it is indispensable., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The incidence of infection by the dengue virus (DENV) has grown dramatically, reaching 128 countries in tropical and subtropical regions worldwide, with a pattern of hyper-endemicity. DENV is a mosquito-borne disease having four serotypes, one or two circulating in epidemic outbreaks. The diagnosis of DENV is challenging mainly due to the circulation of new viruses with remarkable similarities, such as Zika (ZIKV) that may cause fetal microcephaly. DENV affects 390 million people per year, but these numbers may be higher due to the underreported and misclassified cases. Recently, the NS1 nonstructural protein has been described in serum and urine of DENV and ZIKV patients, suggesting its use as a biomarker for screening since a negative NS1 sample confirms the absence of these infections. Herein, a label-free immunosensor comprising an assembled nanostructured thin film of carbon nanotube-ethylenediamine is described. The advantage of in situ electrosynthesis of polymer film is to allow major control of thickness and conductivity, in addition to designing the reactive groups for functionalization. A quartz crystal microbalance system was used to estimate the thickness of the polymeric film obtained. The anti-NS1 monoclonal antibodies were immobilized to carbon nanotubes by covalent linkage, permitting a high stability during measurements. Analytical responses to NS1 were obtained by differential pulse voltammetry (DPV), showing a linear range from 20 to 800 ng mL -1 and reproducibility of 3.0%, with a limit of detection (LOD) of 6.8 ng mL - 1 . This immunosensor was capable of detecting ZIKV and DENV NS1 in spiked urine and real serum in a clinical range.Graphical abstract., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

This work reports the design of a novel plastic antibody for cystatin C (Cys-C), an acute kidney injury biomarker, and its application in point-of-care (PoC) testing. The synthetic antibody was obtained by tailoring a molecularly imprinted polymer (MIP) on a carbon screen-printed electrode (SPE). The MIP was obtained by electropolymerizing pyrrole (Py) with carboxylated Py (Py-COOH) in the presence of Cys-C and multiwall carbon nanotubes (MWCNTs). Cys-C was removed from the molecularly imprinted poly(Py) matrix (MPPy) by urea treatment. As a control, a non-imprinted poly(Py) matrix (NPPy) was obtained by the same procedure, but without Cys-C. The assembly of the MIP material was evaluated in situ by Raman spectroscopy and the binding ability of Cys-C was evaluated by the cyclic voltammetry (CV) and differential pulse voltammetry (DPV) electrochemical techniques. The MIP sensor responses were measured by the DPV anodic peaks obtained in the presence of ferro/ferricyanide. The peak currents decreased linearly from 0.5 to 20.0 ng/mL of Cys-C at each 20 min successive incubation and a limit of detection below 0.5 ng/mL was obtained at pH 6.0. The MPPy/SPE was used to analyze Cys-C in spiked serum samples, showing recoveries <3%. This device showed promising features in terms of simplicity, cost and sensitivity for acute kidney injury diagnosis at the point of care.

Zika virus (ZIKV) is a mosquito-borne infection, predominant in tropical and subtropical regions causing international concern due to the ZIKV disease having been associated with congenital disabilities, especially microcephaly and other congenital abnormalities in the fetus and newborns. Development of strategies that minimize the devastating impact by monitoring and preventing ZIKV transmission through sexual intercourse, especially in pregnant women, since no vaccine is yet available for the prevention or treatment, is critically important. ZIKV infection is generally asymptomatic and cross-reactivity with dengue virus (DENV) is a global concern. An innovative screen-printed electrode (SPE) was developed for amperometric detection of the non-structural protein (NS2B) of ZIKV by exploring the intrinsic redox catalytic activity of Prussian blue (PB), incorporated into a carbon nanotube-polypyrrole composite. Thus, this immunosensor has the advantage of electrochemical detection without adding any redox-probe solution (probe-less detection), allowing a point-of-care diagnosis. It was responsive to serum samples of only ZIKV positive patients and non-responsive to negative ZIKV patients, even if the sample was DENV positive, indicating a possible differential diagnosis between them by NS2B. All samples used here were confirmed by CDC protocols, and immunosensor responses were also checked in the supernatant of C6/36 and in Vero cell cultures infected with ZIKV.

In post-operative scenarios of arterial graft surgeries to bypass coronary artery stenosis, fluid dynamics plays a crucial role. Problems such as intimal hyperplasia have been related to fluid dynamics and wall shear stresses near the graft junction. This study focused on the question of the use of Newtonian and non-Newtonian models to represent blood in this type of problem in order to capture important flow features, as well as an analysis of the performance of geometry from the view of Constructive Theory. The objective of this study was to investigate the effects rheology on the steady-state flow and on the performance of a system consisting of an idealized version of a partially obstructed coronary artery and bypass graft. The Constructal Design Method was employed with two degrees of freedom: the ratio between bypass and artery diameters and the junction angle at the bypass inlet. The flow problem was solved numerically using the Finite Volume Method with blood modeled employing the Carreau equation for viscosity. The Computational Fluid Dynamics model associated with the Sparse Grid method generated eighteen response surfaces, each representing a severe stenosis degree of 75% for specific combinations of rheological parameters, dimensionless viscosity ratio, Carreau number and flow index at two distinct Reynolds numbers of 150 and 250. There was a considerable dependence of the pressure drop on rheological parameters. For the two Reynolds numbers studied, the Newtonian case presented the lowest value of the dimensionless pressure drop, suggesting that the choice of applying Newtonian blood may underestimate the value of pressure drop in the system by about 12.4% (Re =150) and 7.8% (Re = 250). Even so, results demonstrated that non-Newtonian rheological parameters did not influence either the shape of the response surfaces or the optimum bypass geometry, which consisted of a diameter ratio of 1 and junction angle of 30°. However, the viscosity ratio and the flow index had the greatest impact on pressure drop, recirculation zones and wall shear stress. Rheological parameters also affected the recirculation zones downstream of stenosis, where intimal hyperplasia is more prevalent. Newtonian and most non-Newtonian results had similar wall shear stresses, except for the non-Newtonian case with high viscosity ratio. In the view of Constructal Design, the geometry of best performance was independent of the rheological model. However, rheology played an important role on pressure drop and flow dynamics, allowing the prediction of recirculation zones that were not captured by a Newtonian model., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

An interdigitated immunosensor for Cystatin C detection based on polypyrrole/carbon nanotube electrochemical capacitor is described. Cystatin C (CysC) is powerful biomarker for early acute renal failure and one predictive for cardiovascular risk, sepsis, cancer and death. Recently, electrochemical immunosensors based on interdigitated electrodes (IDE) have been successfully focused on development of point-of-care testing, due to their miniaturization facilities and higher sensitivity as compared with the screen-printed electrochemical sensing. Herein, a polypyrrole/carbon nanotube nanoyhibrid film was grafted on two gold fingers by electropolymerization obtaining a supercapacitor. Anti-CysC antibodies were immobilized on the IDE by covalent entrapment via ethylenediamine bifunctional agent, followed by glycine blocking in acid and alkaline medium. Under low frequency, capacitive effect of antigen-antibody interaction were observed by double layer capacitance, and analytical responses of this IDE immunosensor to CysC serum were obtained by changes on phase angle a linear range up to 300 ng/mL. The cutoff was calculated for serum samples showing a total reducing of non-specific binding at approximately 28 ng/mL CysC. This immunosensor based on interdigitated electrode (IDE) is a potential tools as portable device,with possibility to use as a practical and rapid test for CysC diagnostic in samples of serum., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Dengue is a viral disease that represents a significant threat to global public health since billions of people are now at risk of infection by this mosquito-borne virus. The implementation of extensive screening tests is indispensable to control this disease, and the Dengue virus non-structural protein 1 (NS1) is a promising antigen for the serological diagnosis of dengue fever. Plant-based systems can be a safe and cost-effective alternative for the production of dengue virus antigens. In this work, two strategies to produce the dengue NS1 protein in Nicotiana benthamiana leaves were evaluated: Targeting NS1 to five different subcellular compartments to assess the best subcellular organelle for the expression and accumulation of NS1, and the addition of elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags to NS1. The transiently expressed proteins in N. benthamiana were quantified by Western blot analysis. The NS1 fused to ELP and targeted to the ER (NS1 ELP-ER) showed the highest yield (445 mg/kg), approximately a forty-fold increase in accumulation levels compared to the non-fused protein (NS1-ER), representing the first example of transient expression of DENV NS1 in plant. We also demonstrated that NS1 ELP-ER was successfully recognized by a monoclonal anti-dengue virus NS1 glycoprotein antibody, and by sera from dengue virus-infected patients. Interestingly, it was found that transient production of NS1-ER and NS1 ELP-ER using vacuum infiltration of whole plants, which is easier to scale up, rather than syringe infiltration of leaves, greatly improved the accumulation of NS1 proteins. The generated plant made NS1, even without extensive purification, showed potential to be used for the development of the NS1 diagnostic tests in resource-limited areas where dengue is endemic., (Copyright © 2020 Marques, Silva, Dutra, Florean, Menassa and Guedes.)

A novel electrochemical sensor with inherent redox activity mediated by ferrocene for Cystatin C (CysC), an early kidney failure biomarker, is described. The current response was mediated by graphene oxide-ferrocene nanofilm with redox-activity coming from electroactive species surface-confined. Anti-CysC antibodies were immobilized by their Fc portions on the drop-casting polyethyleneimine (PEI) film for improving the sensitivity and reproducibility. Stepwise modifications of the nanostructured surface were characterized by electrochemical techniques, FT-IR and AFM. FT-IR confirmed the formation of the Fc-GO nanocomposite and PEI deposition on the electrode surface. The AFM micrographs confirmed a nanometric film of Fc-GO and PEI. The sensor platform showed a response from 0.1 to 1000 ng/mL and lower limit of detection (LOD) of 0.03 ng/mL of CysC, with good accuracy, specificity and it was successfully applied for CysC detection. Advantages of this immunosensor include rapid testing with minimal steps by the simple use of an intrinsic redox probe, working in a reduction potential, which avoids potential interferences. This proposal attempts to circumvent amperometric detection limitations and provides a promising candidate for future point-of-care diagnostics without redox probe additional solutions for measurements., (Copyright © 2019 Elsevier B.V. All rights reserved.)

An electrochemical immunosensor devoted the core hepatitis B antibody detection, based on polytyramine (PTy) and carbon nanotubes (CNT) composite was developed. The antibody interactions with immobilized antigens were detected by reduction on the electron transfer from ionic species coming from reactive amine groups of PTy. The synthesis in acid medium of PTy-CNT composite favorite a great amount of NH 3 + ionic species, forming a nanocomposite with high catalytic activity on the electrode surface. As proof-of-concept, antibodies against the core hepatitis B virus were label-free and reagentless electrochemically detected by square wave voltammetry (SWV) through decrease on cathodic peaks. It was recently reported that hepatitis B core antigen antibodies (anti-HBc) is a powerful biomarker for hepatitis B virus (HBV) infection, being more specific than HBsAg due to the possibility of detecting the occult HBV infections. The nanostructured film was characterized by atomic force microscopy and electrochemical techniques. This immunosensor showed linear responses from 1.0 to 5.0 ng mL -1 and a limit of detection of 0.89 ng mL -1 anti-HBc. It was also tested in assays with negative and positive blood samples using 0.1 M KCl as electrolyte support on readings showing specific responses. This easy reagentless detection platform, showing a remarkable potential to development of bolder and simpler HBV assay for screening of blood bags, in attempting to circumvent point-of-care testing limitations., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Lymphatic filariasis (LF) is a neglected tropical disease transmitted by mosquitoes and the second cause of permanent disability leading to a significant morbidity and mortality rate. Previously, we have identified epitopes of the filarial abundant larval transcript-2 (ALT-2) protein using a microarray mapping. In this study, one of the epitopes (Wb/ALT2-A5) was used to construct an electrochemical immunosensor. Electrochemical technique of cyclic voltammetry was performed for detecting the signal generated by the interaction between the (Wb/ALT2-A5) peptide and circulating antibodies of serum human samples. (Wb/ALT2-A5) epitope antigens were successfully immobilized on the working electrode of a screen-printed carbon electrode (SPCE) by their amine groups via chitosan film by coupling with glutaraldehyde as crosslinker. After the sensor ready, a pool of human sera infected with Wuchereria bancrofti was added to its surface. Electrochemical responses were generated by applying a potential of - 0.6 to 0.6 V, scan rate of 0.025 V/s. A detection limit of 5.0 µg mL -1 for the synthetic peptides (Wb/ALT2-A5) and 0.002 µg mL -1 for human serum, with a sensitivity of 1.86 µA. The performance of this assay was successfully tested in human serum samples from infected and healthy patients. Thus, this proposed immunosensor, which is able to identify circulating antibodies, can be applied to the diagnosis of the W. bancrofti parasitic disease., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The research for new biomarkers of cancer has studied the role of fetuin glycoprotein on the metastatic disease diagnosis. Cratylia mollis is a lectin with high finity to fetuin, and used here to differentiate prostate cancer and benign prostatic hyperplasia. A label-free electrochemical nanosensor based on assembled carboxylated carbon nanotubes (COOH-CNTs) and poly-L-lysine (PLL) film was developed and applied to serum samples of prostate cancer positive for Gleason score. The electrode analytical response to fetuin in PBS samples, obtained by square wave voltammetry, exhibited a linear range from 0.5 to 25µgmL(-1), with a high correlation coefficient (r=0.994, p<0.001) and low limit of detection (0.017µgmL(-1)). The lectin nanoelectrode showed a good repeatability (1.24% RSD) and reproducibility (4.24% RSD). A pool of serum samples from prostate cancer patients with known the Gleason score were tested showing a significant statistically correlation. Thus, the lectin nanoelectrode was able to distinguish the degree of staging prostate cancer, providing the diagnostic differentiation of benign and malign hyperplasia. To the best of our knowledge, it is the first biosensor for this application using a lectin., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A nano-molecularly imprinted polymer (N-MIP) assembled on a screen-printed electrode for the cardiac troponin T (cTnT) was developed. The biomimetic surface was obtained by a co-polymer matrix assembled on the reduced graphene oxide (RGO) electrode surface. The cTnT active sites were engineered using pyrrole and carboxylated pyrrole that was one-step electropolymerized jointly with cTnT by cyclic voltammetry. The stepwise preparation of the biomimetic surface was characterized by cyclic and differential pulse voltammetries using the ferrocyanide/ferricyanide as redox probe. Structural and morphological characterization was also performed. The optimal relation of pyrrole and pyrrole-3-acid carboxylic to perform the cTnT biomimetic nanosurface was obtained at 1:5 ratio. The analytical performance of cTnT N-MIP performed by differential pulse voltammetry showed a linear range from 0.01 to 0.1 ngmL(-1) (r=0.995, p«0.01), with a very low limit of detection (0.006 ngmL(-1)). The synergic effect of conductive polymer and graphene forming 3D structures of reactive sites resulted in a N-MIP with excellent affinity to cTnT binding (KD=7.3 10(-13) molL(-1)). The N-MIP proposed is based on a simple method of antibody obtaining with a large potential for point-of-care testing applications., (Copyright © 2015 Elsevier B.V. All rights reserved.)

An electrochemical immunosensor developed for detection of antibodies to hepatitis B core protein (anti-HBc) is described. Anti-HBc is the earliest serological marker from hepatitis B virus (HBV) infection, remaining all life after contact with virus, being considered the most important marker for uses in screening of blood bank. A nanohybrid surface assembled onto a glassy carbon electrode consisting of amino carbon nanotubes recovered by hyaluronic acid was used as sensing platform to detect the anti-HBc. All the steps of electrode surface modification were characterized by Scanning Electronic Microscopy and extensively evaluated by electrochemical techniques. The electrode response was measured by direct anti-HBc antigen interactions by square wave voltammetry, dispensing uses of label or chemical mediators. Under optimal conditions, the anodic peak current which was proportional to the anti-HBs concentration. The immunosensor response was linear toward anti-HBc in concentrations up to 6 ng mL(-1), with a detection limit of 0.03 ng mL(-1). The linear range achieved was according to clinical level, indicating the immunosensor as promising tool for use as a criterion for blood bag disposal. The enhancement of the hyaluronic acid by carbon nanotube promoted an increase of charge electron transfer, besides a stable platform for HBc., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A thiophene-modified screen printed electrode (SPE) for detection of the Dengue virus non-structural protein 1 (NS1), an important marker for acute phase diagnosis, is described. A sulfur-containing heterocyclic compound, the thiophene was incorporated to a carbon ink to prepare reproducible screen printed electrodes. After cured, the thiophene SPE was coated by gold nanoparticles conjugated to Protein A to form a nanostrutured surface. The Anti-NS1 antibodies immobilized via their Fc portions via Protein A, leaving their antigen specific sites free circumventing the problem of a random antibodies immobilization. Amperometric responses to the NS1 protein of dengue virus were obtained by cyclic voltammetries performed in presence of ferrocyanide/ferricyanide as redox probe. The calibration curve of immunosensor showed a linear response from 0.04 µg mL(-1) to 0.6 µg mL(-1) of NS1 with a good linear correlation (r=0.991, p<0.05). The detection limit (0.015 µg mL(-1) NS1) was lower than conventional analytical methods. In this work, thiophene monomers incorporated in the carbon ink enhanced the electroanalytical properties of the SPEs, increasing their reproducibility and sensitivity. This point-of-care testing represents a great potential for use in epidemic situations, facilitating the early diagnosis in acute phase of dengue virus., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Diseases such as leishmaniases are important causes of morbidity and mortality in Brazil, and their diagnoses need to be improved. The use of monoclonal antibodies has ensured high specificity to immunodiagnosis. The development of an immunosensor, coupling a monoclonal antibody to a bioelectronic device capable of quickly detecting Leishmania sp. antigens both qualitatively and quantitatively, is a promising alternative for the diagnosis of leishmaniasis due to its high specificity, low cost, and portability, compared with conventional methods. The present work was aimed at developing an immunosensor-based assay for detecting Leishmania infantum antigens in tissues of infected hosts. Four hybridomas producing monoclonal antibodies against L. infantum had their specificity confirmed by enzyme-linked immunosorbent assay. These antibodies were immobilized on a gold surface, covered with a thin film of 2-aminoethanethiol (cysteamine) and glutaraldehyde, blocked with glycine, and placed into contact with extracts of L. infantum -infected and noninfected control hamster spleens. The assay was able to detect 1.8 × 10(4) amastigotes/g of infected tissue. These results demonstrated that this assay may be useful for quantifying L. infantum amastigotes in organs of experimental animals for studies on pathogenesis and immunity and that it is a promising tool for the development of a diagnostic method, based on antigen detection, of human and dog visceral leishmaniasis.

A simple amino-functionalization method for carbon nanotubes and its application in an electrochemical immunosensor for detection of the human cardiac troponin T are described. Amino-functionalized carbon nanotubes allow oriented antibodies immobilization via their Fc regions, improving the performance of an immunosensor. Herein multiwalled carbon nanotubes were amino-functionalized by using the ethylenediamine reagent and assays were designed by fractional factorial study associated with Doehlert matrix. Structural modifications in the carbon nanotubes were confirmed by Fourier transform infrared spectroscopy. After amino-functionalization the carbon nanotubes were attached to screen-printed carbon electrode and a sandwich-type immunoassay was performed for measuring the cardiac troponin T. The electrochemical measurements were obtained through hydrogen peroxide reaction with peroxidase conjugated to the secondary antibody. Under optimal conditions, troponin T immunosensor was evaluated in serum samples, which showed a broad linear range (0.02 to 0.32 ng mL(-1)) and a low limit of detection, 0.016 ng mL(-1). This amino platform can be properly used as clinical tool for cardiac troponin T detection in the acute myocardial infarction diagnosis.

Label-free immunosensor based on amine-functionalized carbon nanotubes screen-printed electrode is described for detection of the cardiac troponin T, an important marker of acute myocardial infarction. The disposable sensor was fabricated by tightly squeezing an adhesive carbon ink containing carbon nanotubes onto a polyethylene terephthalate substrate forming a thin film. The use of carbon nanotubes increased the reproducibility and stability of the sensor, and the amine groups permitted nonrandom immobilization of antibodies against cardiac troponin T. Amperometric responses were obtained by differential pulse voltammetry in presence of a ferrocyanide/ferricyanide redox probe after troponin T incubation. The calibration curve indicated a linear response of troponin T between 0.0025 ng mL(-1) and 0.5 ng mL(-1), with a good correlation coefficient (r=0.995; p<0.0001, n=7). The limit of detection (0.0035 ng mL(-1) cardiac troponin T) was lower than any previously described by immunosensors and was comparable with conventional analytical methods. The high reproducibility and clinical range obtained using this immunosensor support its utility as a potential tool for point-of-care acute myocardial infarction diagnostic testing., (© 2013 Elsevier B.V. All rights reserved.)

Dengue fever is a viral disease that affects millions of people worldwide. Specific tests for dengue are not usually performed due to high costs, complicated procedures and, in some cases, long time to yield a result. For widespread use of specific tests to be possible, fast, reliable and fairly simple methods are needed. In this paper, we present a new dengue diagnostic method for the acute phase of the infection. The method proposed uses an all-optical fiber sensor based on Localized Surface Plasmon Resonance (LSPR) and specular reflection from gold nanoparticles (AuNPs). Dengue anti-NS1 antibody was immobilized on AuNPs deposited on the endface of a standard multimode fiber (62.5 µm/125 µm). The sensor is able to detect NS1 antigen at different concentrations, with limit of quantification estimated to be 0.074 μg/ml = 1.54 nM. These results indicate that the sensor could potentially be used for dengue diagnosis in the acute phase of the infection.

An immunosensor for the non-structural protein 1 (NS1) of the dengue virus based on carbon nanotube-screen printed electrodes (CNT-SPE) was successfully developed. A homogeneous mixture containing carboxylated carbon nanotubes was dispersed in carbon ink to prepare a screen printed working electrode. Anti-NS1 antibodies were covalently linked to CNT-SPE by an ethylenediamine film strategy. Amperometrical responses were generated at -0.5 V vs. Ag/AgCl by hydrogen peroxide reaction with peroxidase (HRP) conjugated to the anti-NS1. An excellent detection limit (in the order of 12 ng mL(-1)) and a sensitivity of 85.59 μA mM(-1)cm(-2) were achieved permitting dengue diagnostic according to the clinical range required. The matrix effect, as well as the performance of the assays, was successfully evaluated using spiked blood serum sample obtaining excellent recovery values in the results. Carbon nanotubes incorporated to the carbon ink improved the reproducibility and sensitivity of the CNT-SPE immunosensor. This point-of-care approach represents a great potential value for use in epidemic situations and can facilitate the early screening of patients in acute phase of dengue virus., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This study was carried out to develop a simple and inexpensive method for detection of Human papillomavirus (HPV 18) based on irreversible immobilization of ultra specific primer on silanized glass slides. This method is revealed by Blue Green Loading Dye I (LGC) and compared with conventional polymerase chain reaction (PCR) for endocervical samples. The new method was tested in 40 DNA samples with precancer uterine lesions of women treated in Hospital of Recife PE, Brazil. DNA samples were extracted using Wizard Genomic DNA Purification Kit according to manufacturer's instructions. The samples were tested for HPV 18 by conventional PCR (PCRc) and the products visualized on 1.2% agarose gel, with LGC under UV 260 nm. After that the positive and negative samples to HPV 18 were tested by immobilization method and the results visualized with LGC under UV 260 nm. Both PCRc and immobilization method showed high degree of correlation (95%), whereas comparison between PCRc and immobilization method showed good correlation (100%). PCRc is widely known for detection of HPV because of its high sensitivity and efficiency, but due to high cost it is not yet standardized for use in public health laboratories. In our study, the single-stranded DNA immobilized method on a glass slide was effective in screening for HPV revealed by Blue Green and may be an alternative method for diagnosis of HPV once it offers a fast, and easy handling., (© 2013 Wiley Periodicals, Inc.)

Oil from the seed of the castor plant (Ricinus communis L.) is an important commodity for a number of industries, ranging from pharmaceuticals to renewable energy resources. However, the seed and subsequent seed meal contain ricin (RCA60), a potent cytotoxin, making it an unusable product for animal feed. In order to investigate the efficiency of reducing the toxicity of the seed meal, a biosensor is proposed by exploring the lectin-carbohydrate binding. A gold electrode was assembled with a film of Xyloglucan (XG) extracted from Hymenaea courbaril L. The analytical response to RCA60 was obtained using a polyclonal antibody against RCA60 conjugated to peroxidase. The current responses were generated by reaction with H2O2 and amplified with hydroquinone as chemical mediator. Voltammetric studies showed that the XG film was tightly bound to the gold electrode. This biosensor allows discriminate lectins in native and denatured forms. The limit of detection of native RCA60 was 2.1 μg mL(-1). This proposed biosensor showed to be a potential and accurate method for toxicity assessment of the ricin in castor seed meal by simple polysaccharide film-electrode strategy., (Copyright © 2012. Published by Elsevier Ltd.)

A chitosan-modified carbon fiber electrode (CFE) for dengue virus envelope protein (DENV) was developed. Antibodies against DENV were covalently immobilized on the chitosan (CHIT) matrix after activation with sodium periodate. Cyclic voltammetries and scanning electron microscopies analysis were performed to monitor steps involved in the CFE surface modification. Amperometric response of the competitive immunoassays was generated by hydrogen peroxide reaction with the peroxidase conjugated to DENV and 2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as mediator. The immunosensor showed a lower limit of detection for DENV (0.94 ng mL(-1)) than previously described and a linear range from 1.0 to 175 ng mL(-1), in concentration levels clinically relevant for dengue virus diagnosis. The intra- and inter-assay were respectively 5.8% and 3.6%. The unique and simple design of this immunoassay format provides an economical alternative for the manufacture of other sensitive sensors., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Aedes aegypti and Aedes albopictus perform an important role in the transmission of the dengue virus to human populations, particularly in the tropical and subtropical regions of the world. Despite a lack of understanding in relation to the maintenance of the dengue virus in nature during interepidemic periods, the vertical transmission of the dengue virus in populations of A. aegypti and A. albopictus appears to be of significance in relation to the urban scenario of Fortaleza., Methods: From March 2007 to July 2009 collections of larvae and pupae of Aedes spp were carried out in 40 neighborhoods of Fortaleza. The collections yielded 3,417 (91%) A. aegypti mosquitoes and 336 (9%) A. albopictus mosquitoes. Only pools containing females, randomly chosen, were submitted to the following tests indirect immunofluorescence (virus isolation), RT-PCR/nested-PCR and nucleotide sequencing at the C-prM junction of the dengue virus genome., Results: The tests on pool 34 (35 A. albopictus mosquitoes) revealed with presence of DENV-3, pool 35 (50 A. aegypti mosquitoes) was found to be infected with DENV-2, while pool 49 (41 A. albopictus mosquitoes) revealed the simultaneous presence of DENV-2 and DENV-3. Based on the results obtained, there was a minimum infection rate of 0.5 for A. aegypti and 9.4 for A. albopictus. The fragments of 192 bp and 152 bp related to DENV-3, obtained from pools 34 and 49, was registered in GenBank with the access codes HM130699 and JF261696, respectively., Conclusions: This study recorded the first natural evidence of the vertical transmission of the dengue virus in populations of A. aegypti and A. albopictus collected in Fortaleza, Ceará State, Brazil, opening a discuss on the epidemiological significance of this mechanism of viral transmission in the local scenario, particularly with respect to the maintenance of these viruses in nature during interepidemic periods.

The American visceral leishmaniasis is an important cause of morbidity and mortality in Brazil for both humans and dogs. Attempts to make a diagnosis of this disease need to be improved, especially in endemic areas, and in the tracking and screening of asymptomatic dogs, which are their main host in urban areas. A quartz crystal microbalance immunosensor for the diagnosis of the canine visceral leishmaniasis using a recombinant antigen of Leishmania chagasi (rLci2B-NH6) was developed. The rLci2B-NH6 was tightly immobilized on a quartz crystal gold electrode by self-assembled monolayer based on short-chain length thiol. The strategy was the use of the antigen-histidine tail covalently linked to glutaraldehyde performing a Schift base which permits a major exposure of epitopes and a reduced steric hindrance. The immunosensor showed good results regarding sensitivity and reproducibility, being able to distinguish positive and negative canine serum for L. chagasi. Furthermore, the immunosensor can be reused through exposure to sodium dodecyl sulfate solution, which promotes the dissociation of antigen-antibody binding, restoring the sensor surface with immobilized biologically active antigens for further analysis.

A piezoelectric immunosensor based on gold nanoparticles (AuNPs) co-immobilized on a dithiol-modified surface is proposed for detection of human cardiac troponin T (TnT). Anti-human troponin T (anti-TnT) antibodies were covalently immobilized on the nanostructured electrode surface by thiol-aldehyde linkages. In a homogeneous bulk solution, TnT was captured by anti-TnT immobilized on the QCM electrode. Cyclic voltammetry studies were used to characterize the AuNPs layer on the electrode surface and the anti-TnT immobilization steps. The QCM-flow immunosensor exhibited good reliability, measuring concentrations of TnT from 0.003 to 0.5 ng mL(-1) in human serum with high linearity (r = 0.989; p < 0.01). The immunosensor exhibited a 7% coefficient of variation and 0.0015 ng mL(-1) limit of detection, indicating a high reproducibility and sensitivity. The proposed QCM nanostructured immunosensor is easy to use and has promising potential in the diagnosis of acute myocardial infarction due to its speed and high sensitivity.

Screen-printed electrodes (SPE) have been widely used in the design of disposable sensors bringing advances in the use of electrochemical immunosensors for in field-clinical analysis. In this work, streptavidin polystyrene microspheres were incorporated to the electrode surface of SPEs in order to increase the analytical response of the cardiac troponin T (cTnT), a specific biomarker for the acute myocardial infarction diagnosis. The precise calculation of the stoichiometric streptavidin-biotin ratio [1:4] allowed the increase of sensitivity and stability of the immunosensor response to the cTnT analyte. The surface of the immunosensor was characterized by scanning electron microscopy and cyclic voltammetry. It was observed that the use of streptavidin microspheres significantly increased the analytical sensitivity of the electrode in 8.5 times, showing a curve with a linear response range between 0.1 and 10 ngmL(-1) of cTnT and a detection limit of 0.2 ngmL(-1). The proposed SPE showed ease preparation and high sensitivity allowing the detection of cTnT in the range of clinical levels. The new device coupled with a portable electrochemical analyzer shows great promise for point-of-care quantitative testing of necrosis cardiac proteins., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Purpose: The impact of weight on quality of life (IWQOL-Lite) is a self-administered instrument that assesses quality of life in obesity. It is composed of 31 items and five domains (physical function, self-esteem, sexual life, public distress, and work). The aim of the present study was to evaluate reliability (test-retest), internal structure, construct validity and discriminant validity., Methods: IWQOL-Lite scores were obtained from 89 people in a Weight Watchers institution (clinical sample) and 156 community volunteers (community sample). The participants were selected based on the same criteria: gender (female) and age (> 24 years), except for BMI. The community sample also completed the SF-36, a generic measure of health-related quality of life., Results: The Brazilian IWQOL-Lite demonstrated good test-retest reliability, internal consistency, discriminative validity and convergent validity., Conclusions: The IWQOL-Lite is the first specific instrument validated in Brazil for assessing quality of life in obesity. The results reveal that IWQOL-Lite is an instrument with good psychometric properties. Nevertheless, in some cases results were dissimilar to those reported in earlier studies using the original American English.

Introduction: Since the registration of Aedes albopictus in the municipality of Fortaleza in 2005, it has shown rapid dispersion. A study aiming to identify its occurrence areas, breeding sites and associations with Aedes aegypti and other culicids was carried out during 2008., Methods: Between January and July 2008, samples of immature forms of culicids were gathered from properties located in districts of Fortaleza. Certain features of the breeding sites were highlighted, such as location (indoors or outdoors), presence of covering (protection against impact of sunlight and rain), water turbidity (water clear or not clear), constituent material, volume, height above ground level and simultaneous presence of different species of culicids in the same breeding site., Results: Indoor location was an important factor for the breeding sites for Aedes albopictus [PR = 0.52; 95% CI: 0.33-0.81]. On the other hand, water turbidity and reservoir covering were not shown to be differentiators regarding infestation [p > 0.05]. Absence of water turbidity was significant for Aedes aegypti infestation in breeding sites [PR = 1.14; 95% CI: 1.06-1.22]., Conclusions: Absence of one of the species from the breeding sites enhanced the infestation by another species. Breeding sites that had not been infested by Aedes albopictus had a prevalence of infestation by Aedes aegypti that was 2.05 times greater [95% CI: 1.72-2.44]. There was no significant association between volume and height of the breeding sites and infestation by both species. Both species were found to be dispersed throughout the municipality, occupying a wide diversity of breeding sites. However, a slight physical separation could be identified, with higher infestation with Aedes albopictus outdoors.

A simplified measurement scheme and device structure aiming at developing a low cost, label-free, point-of-care capacitive biosensor were investigated. The detection principle is the increase of low frequency capacitance between two planar Al electrodes observed after antibody-antigen interaction. The electrodes, deposited on oxidized Si wafers, were covered with an antibody layer, with and without using self-assembled thiol monolayer. Immunoglobulin G (IgG) and cardiac troponin T (TnT) were used as analytes to asses this proposal. The device was able to detect successfully TnT levels in the range 0.07 to 6.83ng/mL in human serum from patients with cardiac diseases and in the range 0.01ng/mL to 5ng/mL for TnT in phosphate buffer saline. An equivalent circuit model able to reproduce the general behavior of experimental capacitance versus frequency curves was presented. The investigated features that have potential to reduce costs and simplify measurements were: use of single, low frequency (1kHz) measurement signal, within the range of low cost portable capacitance meters; employment of a lower cost electrode material, aluminum, instead of gold electrodes; and use of simple and miniaturized planar two-electrodes arrangement, thus making a portable system for point-of-care applications.

The cardiac troponin T (cTnT) is specific biomarker important for trials of acute myocardial infarctions (AMI). In this paper, a SPR sensor in real time to detect the biomarker was developed on a commercially available surface plasmon resonance AUTOLAB SPIRIT. The cTnT receptor molecule was covalently immobilized on a gold substrate via a self-assembled monolayer (SAM) of thiols by using cysteamine-coupling chemistry. This biosensor presented a linear response range for cTnT between 0.05 and 4.5 ng/mL (r=0.997, p<<0.01) with a good reproducibility (CV=4.4%). The effect of the cysteamine (CYS) concentrations on the SAM coated gold sensor was studied as a function of the amount of the immobilized cTnT monoclonal antibodies. Analysis using serum samples undiluted was carried out at room temperature showing a well agreement with the ECLIA methods and the sensor surface could be regenerated by using a solution of 1% (w/v) sodium dodecyl sulphate (SDS) without losing the sensor immunoreactivity. These studies open new perspectives of using SAM to develop regenerable immunosensor with a good reproducibility allowing its use in the clinical applications.

Background: The cardiac troponin T (TnT) is a cardiospecific, highly sensitive marker for myocardial damage and is immediately released to bloodstream during the acute myocardial infarction (AMI). Surface plasmon resonance (SPR) immunosensor was developed for the quick detection of human cTnT in real time., Methods: An SPR sensor streptavidin terminated self-assembled monolayer, which was used to binding biotinylated anti troponin T monoclonal antibodies, was developed., Results: The cTnT was determined from successive injections with a linear range from 0.03 up to 6.5 ng/ml. The detection limit was 0.01 ng/ml corresponding to a resonant angle change of 1.28 millidegrees. The system presented a good repeatability with 3.4% of variation between run after regeneration of the coated surface with a solution of 1% SDS. The effects of blocking of non-specific adsorption using different solutions were studied. It was possible to measure the cTnT without dilution of the human serum with good specificity and reproducibility., Conclusion: This sensor is practical and offer quick response in interval of 800 s.

The electrochemical potentials for free or immobilized Cratylia mollis seed lectin (Cra) were obtained through potentiostatic techniques. A saline solution was used as support to control the charge distribution between saturated calomel electrode and platinum electrode (working electrode). The electrochemical potential to free Cra was determined at the following concentrations: 0.6, 0.9 and 1.0 mg/ml in an aerated environment under different temperatures (5, 10 and 20 degrees C). The best electrochemical potential was obtained with 1.0 mg/ml, at 5 and 10 degrees C, 87 and 102 mV, respectively. Electrochemical potential to Cra immobilized on glass beads activated with 3-aminopropyltriethoxysilane described a linear behavior in relation to the increase in glucose concentration. The development of techniques to define interface electrical parameters will be able to give information about charged groups adsorbed to electrode surface revealing interactions particularly in biological systems.

One the most important aspects of a biosensor is related to immobilization and maintenance of specific reference compounds on sensing surfaces. A method for the immobilization of polysaccharides to a silicon oxide surface intended for Surface Acoustical Waves (SAW) sensors is described. Silicon oxide is a hydrophobic inorganic support used for the fabrication of many electronic devices. The pneumococcal polysaccharide (PPS) vaccine is immobilized via Protein A after pre-treatment of the surface with hydrochloric acid. The effects of non-specific binding are discussed. The results indicate that the immobilization of PPS via Protein A increases the sensitivity of detecting Streptococcus pneumoniae antibodies in human sera and offers greater reproducibility of response compared with ELISA methods. The principles of this technique are simple and are applicable to the immobilization of many capsular polysaccharides.

Report a case of a patient suffering from acute pancreatitis with renal failure, who needed peritoneal dialysis and developed acute hydrothorax as a complication of the procedure. The diagnosis was made by the high levels of glucose on pleural effusion (455 mg/dL) and glycemia of 81 mg/dL). This complication probably occurs because of a pathological diaphragmatic defect, communicating peritoneal and pleural cavities. The treatment consists of prompt interruption of peritoneal dialysis with improvement of symptomatology.