Development of an Electrochemical Immunosensor for Specific Detection of Visceral Leishmaniasis Using Gold-Modified Screen-Printed Carbon Electrodes

Visceral leishmaniasis is a reemerging neglected tropical disease with limitations for its diagnosis, including low concentration of antibodies in the serum of asymptomatic patients and cross-reactions. In this context, this work proposes an electrochemical immunosensor for the diagnosis of visceral leishmaniasis in a more sensitive way that is capable of avoiding cross-reaction with Chagas disease (CD). Crude Leishmania infantum antigens tested in the enzyme-linked immunosorbent assay (ELISA) were methodologically standardized to best engage to the sensor. The antibodies anti-Trypanosoma cruzi and anti-Leishmania sp. Present in serum from patients with diverse types of CD or leishmaniasis were chosen. A screen-printed carbon electrode modified with gold nanoparticles was the best platform to guarantee effective adsorption of all antigens so that the epitope of specific recognition for leishmaniasis occurred efficiently and without cross-reaction with the evaluated CD. The current peaks reduced linearly after the recognition, and still were able to notice the discrimination between different kinds of diseases (digestive, cardiac, undetermined Chagas/acute and visceral chronic leishmaniasis). Comparative analyses with ELISA were performed with the same groups, and a low specificity (44%) was verified due to cross-reactions (high number of false positives) on ELISA tests, while the proposed immunosensor presented high selectivity and specificity (100%) without any false positives or false negatives for the serum samples from isolated patients with different types of CD and visceral leishmaniasis. Furthermore, the biosensor was stable for 5 days and presented a detection limit of 200 ng mL−1.

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Development of Highly Sensitive Immunosensor for Clenbuterol Detection by Using Poly(3,4-ethylenedioxythiophene)/Graphene Oxide Modified Screen-Printed Carbon Electrode

Sensors ◽

10.3390/s18124324 ◽

2018 ◽

Vol 18 (12) ◽

pp. 4324 ◽

Cited By ~ 9

Author(s):

Nurul Talib ◽

Faridah Salam ◽

Yusran Sulaiman

Keyword(s):

Graphene Oxide ◽

Limit Of Detection ◽

Food Product ◽

Electrochemical Immunosensor ◽

Growth Promoter ◽

Carbon Electrode ◽

Electrochemical Assay ◽

Sensor Platform ◽

Screen Printed Carbon Electrode ◽

Screen Printed

Clenbuterol (CLB) is an antibiotic and illegal growth promoter drug that has a long half-life and easily remains as residue and contaminates the animal-based food product that leads to various health problems. In this work, electrochemical immunosensor based on poly(3,4-ethylenedioxythiophene)/graphene oxide (PEDOT/GO) modified screen-printed carbon electrode (SPCE) for CLB detection was developed for antibiotic monitoring in a food product. The modification of SPCE with PEDOT/GO as a sensor platform was performed through electropolymerization, while the electrochemical assay was accomplished while using direct competitive format in which the free CLB and clenbuterol-horseradish peroxidase (CLB-HRP) in the solution will compete to form binding with the polyclonal anti-clenbuterol antibody (Ab) immobilized onto the modified electrode surface. A linear standard CLB calibration curve with R2 = 0.9619 and low limit of detection (0.196 ng mL−1) was reported. Analysis of milk samples indicated that this immunosensor was able to detect CLB in real samples and the results that were obtained were comparable with enzyme-linked immunosorbent assays (ELISA).

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Simple Electrochemical Immunosensor for the Detection of Hippuric Acid on the Screen-printed Carbon Electrode Modified Gold Nanoparticles

Journal of the Korean Electrochemical Society ◽

10.5229/jkes.2011.14.1.044 ◽

2011 ◽

Vol 14 (1) ◽

pp. 44-49 ◽

Cited By ~ 2

Author(s):

Young-Bong Choi ◽

Gun-Sik Tae

Keyword(s):

Gold Nanoparticles ◽

Hippuric Acid ◽

Electrochemical Immunosensor ◽

Carbon Electrode ◽

Screen Printed Carbon Electrode ◽

Screen Printed

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Diagnostic application of sensitive and specific phage-exposed epitopes for visceral leishmaniasis and human immunodeficiency virus coinfection

Parasitology ◽

10.1017/s0031182021001505 ◽

2021 ◽

pp. 1-9

Author(s):

Fernanda F. Ramos ◽

Grasiele S. V. Tavares ◽

Fernanda Ludolf ◽

Amanda S. Machado ◽

Thaís T. O. Santos ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Visceral Leishmaniasis ◽

Patient Treatment ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Purpose ◽

Serum Samples ◽

Prognostic Effect ◽

Specific Phage ◽

Hiv Coinfection ◽

Immunodeficiency Virus

Abstract The diagnosis of visceral leishmaniasis (VL) has improved with the search of novel antigens; however, their performance is limited when samples from VL/human immunodeficiency virus (HIV)-coinfected patients are tested. In this context, studies conducted to identify more suitable antigens to detect both VL and VL/HIC coinfection cases should be performed. In the current study, phage display was performed using serum samples from healthy subjects and VL, HIV-infected and VL/HIV-coinfected patients; aiming to identify novel phage-exposed epitopes to be evaluated with this diagnostic purpose. Nine non-repetitive and valid sequences were identified, synthetized and tested as peptides in enzyme-linked immunosorbent assay experiments. Results showed that three (Pep2, Pep3 and Pep4) peptides showed excellent performance to diagnose VL and VL/HIV coinfection, with 100% sensitivity and specificity values. The other peptides showed sensitivity varying from 50.9 to 80.0%, as well as specificity ranging from 60.0 to 95.6%. Pep2, Pep3 and Pep4 also showed a potential prognostic effect, since specific serological reactivity was significantly decreased after patient treatment. Bioinformatics assays indicated that Leishmania trypanothione reductase protein was predicted to contain these three conformational epitopes. In conclusion, data suggest that Pep2, Pep3 and Pep4 could be tested for the diagnosis of VL and VL/HIV coinfection.

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Development of highly sensitive electrochemical immunosensor based on single-walled carbon nanotube modified screen-printed carbon electrode

Materials Chemistry and Physics ◽

10.1016/j.matchemphys.2019.01.068 ◽

2019 ◽

Vol 227 ◽

pp. 123-129 ◽

Cited By ~ 11

Author(s):

Nguyen Xuan Viet ◽

Nguyen Xuan Hoan ◽

Yuzuru Takamura

Keyword(s):

Carbon Nanotube ◽

Electrochemical Immunosensor ◽

Carbon Electrode ◽

Single Walled Carbon Nanotube ◽

Single Walled Carbon ◽

Screen Printed Carbon Electrode ◽

Highly Sensitive ◽

Screen Printed

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Electrochemical Immunosensor Based on Highly Sensitive Amino Functionalized Graphene Nanoplatelets-Modified Screen Printed Carbon Electrode

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.833.171 ◽

2020 ◽

Vol 833 ◽

pp. 171-175

Author(s):

Nurul Azurin Badruzaman ◽

Mohd Azraie Mohd Azmi ◽

Nur Azura Mohd Said

Keyword(s):

Incubation Time ◽

Electrochemical Impedance ◽

Electrochemical Immunosensor ◽

Graphene Nanoplatelets ◽

Carbon Electrode ◽

Functionalized Graphene ◽

Working Electrode ◽

Screen Printed Carbon Electrode ◽

Rabbit Igg ◽

Screen Printed

We presented here the development of an immunosensor based on graphene nanoplatelets-modified screen printed carbon electrode (SPCE) with incorporated rabbit IgG on the amino functionalized surface area. In order to improve sensitivity of working electrode, graphene-nanoplatelets solution was fabricated onto surface carbon working electrode. The effect of different (3-aminopropyl) triethoxysilane (APTES) concentrations (0.125, 0.5, 2 and 8% (v/v)) and incubation time of silanization (30, 60 and 90 min) were studied and compared. An electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to characterize our immunosensor based. It is showed that the optimum APTES concentration which provides higher surface coverage and electron transfer rate was 2% concentration (v/v) at 60 min of incubation time. The modified surface was then evaluated by measuring immobilized rabbit IgG via indirect assay using horseradish peroxidase labelled secondary antibody. The optimum detection immobilized IgG was 0.05 mg/mL. These results indicate the potential for amino functionalized graphene nanoplatelets-modified SPCE in detecting protein biomarkers.

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Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

Clinical and Vaccine Immunology ◽

10.1128/cvi.00583-13 ◽

2013 ◽

Vol 21 (1) ◽

pp. 96-106 ◽

Cited By ~ 18

Author(s):

Lourena E. Costa ◽

Mayara I. S. Lima ◽

Miguel A. Chávez-Fumagalli ◽

Daniel Menezes-Souza ◽

Vivian T. Martins ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Phage Display ◽

Leishmania Infantum ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Phage Display Technology ◽

Predictive Values ◽

Content Type ◽

Phage Elisa

ABSTRACTVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy andTrypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected withLeishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n= 31) compared to those from vaccinated dogs (n= 21), experimentally infected dogs with cross-reactive parasites (n= 23), and healthy controls (n= 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no cross-reactivity withT. cruzi- orEhrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes ofL. infantumantigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs.

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Screen-printed carbon electrode-based electrochemical immunosensor for rapid detection of microalbuminuria

Biosensors and Bioelectronics ◽

10.1016/j.bios.2015.11.002 ◽

2016 ◽

Vol 77 ◽

pp. 1175-1182 ◽

Cited By ~ 27

Author(s):

Jang-Zern Tsai ◽

Ching-Jung Chen ◽

Kalpana Settu ◽

Yu-Feng Lin ◽

Chien-Lung Chen ◽

...

Keyword(s):

Rapid Detection ◽

Electrochemical Immunosensor ◽

Carbon Electrode ◽

Screen Printed Carbon Electrode ◽

Screen Printed

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An electrochemical immunosensor for rapid determination of clenbuterol by using magnetic nanocomposites to modify screen printed carbon electrode based on competitive immunoassay mode

Sensors and Actuators B Chemical ◽

10.1016/j.snb.2013.11.011 ◽

2014 ◽

Vol 192 ◽

pp. 529-535 ◽

Cited By ~ 47

Author(s):

Xin Yang ◽

Feng Wu ◽

Di-Zhao Chen ◽

Hong-Wei Lin

Keyword(s):

Rapid Determination ◽

Electrochemical Immunosensor ◽

Magnetic Nanocomposites ◽

Carbon Electrode ◽

Competitive Immunoassay ◽

Screen Printed Carbon Electrode ◽

Screen Printed

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Diagnosis of Visceral Leishmaniasis by Enzyme-Linked Immunosorbent Assay Using Urine Samples

Clinical and Vaccine Immunology ◽

10.1128/cdli.9.4.789-794.2002 ◽

2002 ◽

Vol 9 (4) ◽

pp. 789-794 ◽

Cited By ~ 3

Author(s):

Mohammad Zahidul Islam ◽

Makoto Itoh ◽

S. M. Shamsuzzaman ◽

Rusella Mirza ◽

Farzana Matin ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Sensitivity And Specificity ◽

Immunosorbent Assay ◽

Leishmania Donovani ◽

Laboratory Diagnosis ◽

Urine Samples ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Controls ◽

Serum Samples ◽

Crude Antigen

ABSTRACT A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone-treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis.

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