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15 pages, 2456 KiB

Open AccessArticle

Cocaine Detection by a Laser-Induced Immunofluorometric Biosensor

by Martin Paul, Robert Tannenberg, Georg Tscheuschner, Marco Ponader and Michael G. Weller

Biosensors 2021, 11(9), 313; https://doi.org/10.3390/bios11090313 - 03 Sep 2021

Cited by 8 | Viewed by 3176

The trafficking of illegal drugs by criminal networks at borders, harbors, or airports is an increasing issue for public health as these routes ensure the main supply of illegal drugs. The prevention of drug smuggling, including the installation of scanners and other analytical [...] Read more.

The trafficking of illegal drugs by criminal networks at borders, harbors, or airports is an increasing issue for public health as these routes ensure the main supply of illegal drugs. The prevention of drug smuggling, including the installation of scanners and other analytical devices to detect small traces of drugs within a reasonable time frame, remains a challenge. The presented immunosensor is based on a monolithic affinity column with a large excess of immobilized hapten, which traps fluorescently labeled antibodies as long as the analyte cocaine is absent. In the presence of the drug, some binding sites of the antibody will be blocked, which leads to an immediate breakthrough of the labeled protein, detectable by highly sensitive laser-induced fluorescence with the help of a Peltier-cooled complementary metal-oxide-semiconductor (CMOS) camera. Liquid handling is performed with high-precision syringe pumps and microfluidic chip-based mixing devices and flow cells. The biosensor achieved limits of detection of 7 ppt (23 pM) of cocaine with a response time of 90 s and a total assay time below 3 min. With surface wipe sampling, the biosensor was able to detect 300 pg of cocaine. This immunosensor belongs to the most sensitive and fastest detectors for cocaine and offers near-continuous analyte measurement. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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14 pages, 1861 KiB

Open AccessCommunication

Nanoribbon-Based Electronic Detection of a Glioma-Associated Circular miRNA

by Yuri D. Ivanov, Kristina A. Malsagova, Vladimir P. Popov, Tatyana O. Pleshakova, Andrey F. Kozlov, Rafael A. Galiullin, Ivan D. Shumov, Svetlana I. Kapustina, Fedor V. Tikhonenko, Vadim S. Ziborov, Alexander Yu. Dolgoborodov, Oleg F. Petrov, Olga A. Gadzhieva, Boris A. Bashiryan, Vadim N. Shimansky, Natalia V. Potoldykova, Dmitry V. Enikeev, Dmitry Yu. Usachev and Alexander I. Archakov

Biosensors 2021, 11(7), 237; https://doi.org/10.3390/bios11070237 - 13 Jul 2021

Cited by 11 | Viewed by 2511

Abstract

Nanoribbon chips, based on “silicon-on-insulator” structures (SOI-NR chips), have been fabricated. These SOI-NR chips, whose surface was sensitized with covalently immobilized oligonucleotide molecular probes (oDNA probes), have been employed for the nanoribbon biosensor-based detection of a circular ribonucleic acid (circRNA) molecular marker of [...] Read more.

Nanoribbon chips, based on “silicon-on-insulator” structures (SOI-NR chips), have been fabricated. These SOI-NR chips, whose surface was sensitized with covalently immobilized oligonucleotide molecular probes (oDNA probes), have been employed for the nanoribbon biosensor-based detection of a circular ribonucleic acid (circRNA) molecular marker of glioma in humans. The nucleotide sequence of the oDNA probes was complimentary to the sequence of the target oDNA. The latter represents a synthetic analogue of a glioma marker—NFIX circular RNA. In this way, the detection of target oDNA molecules in a pure buffer has been performed. The lowest concentration of the target biomolecules, detectable in our experiments, was of the order of ~10⁻¹⁷ M. The SOI-NR sensor chips proposed herein have allowed us to reveal an elevated level of the NFIX circular RNA in the blood of a glioma patient. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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11 pages, 5634 KiB

Open AccessArticle

Development of a Lateral Flow Immunoassay (LFIA) to Screen for the Release of the Endocrine Disruptor Bisphenol A from Polymer Materials and Products

by Anna Raysyan and Rudolf J. Schneider

Biosensors 2021, 11(7), 231; https://doi.org/10.3390/bios11070231 - 11 Jul 2021

Cited by 6 | Viewed by 4304

Abstract

One of the most important chemicals used in the production of polymer plastics and coatings is bisphenol A. However, despite the large number of studies on the toxicity and hormonal activity of BPA, there are still open questions and thus considerable media attention [...] Read more.

One of the most important chemicals used in the production of polymer plastics and coatings is bisphenol A. However, despite the large number of studies on the toxicity and hormonal activity of BPA, there are still open questions and thus considerable media attention regarding BPA toxicity. Hence, it is necessary to develop a sensitive, simple, cost-efficient, specific, portable, and rapid method for monitoring bisphenol A and for high sample throughput and on-site screening analysis. Lateral flow immunoassays have potential as rapid tests for on-site screening. To meet sensitivity criteria, they must be carefully optimized. A latex microparticle-based LFIA for detection of BPA was developed. The sensitivity of the assay was improved by non-contact printing of spot grids as the control and test lines with careful parameter optimization. Results of the test could be visually evaluated within 10 min with a visual cut-off of 10 µg/L (vLOD). Alternatively, photographs were taken, and image analysis performed to set up a calibration, which allowed for a calculated limit of detection (cLOD) of 0.14 µg/L. The method was validated for thermal paper samples against ELISA and LC–MS/MS as reference methods, showing good agreement with both methods. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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13 pages, 3350 KiB

Open AccessArticle

Comparative Study of In Situ Techniques to Enlarge Gold Nanoparticles for Highly Sensitive Lateral Flow Immunoassay of SARS-CoV-2

by Vasily G. Panferov, Nadezhda A. Byzova, Sergey F. Biketov, Anatoly V. Zherdev and Boris B. Dzantiev

Biosensors 2021, 11(7), 229; https://doi.org/10.3390/bios11070229 - 08 Jul 2021

Cited by 15 | Viewed by 3827

Abstract

Three techniques were compared for lowering the limit of detection (LOD) of the lateral flow immunoassay (LFIA) of the receptor-binding domain of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) based on the post-assay in situ enlargement of Au nanoparticles (Au NPs) on a [...] Read more.

Three techniques were compared for lowering the limit of detection (LOD) of the lateral flow immunoassay (LFIA) of the receptor-binding domain of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) based on the post-assay in situ enlargement of Au nanoparticles (Au NPs) on a test strip. Silver enhancement (growth of a silver layer over Au NPs—Au@Ag NPs) and gold enhancement (growth of a gold layer over Au NPs) techniques and the novel technique of galvanic replacement of Ag by Au in Au@Ag NPs causing the formation of Au@Ag-Au NPs were performed. All the enhancements were performed on-site after completion of the conventional LFIA and maintained equipment-free assay. The assays demonstrated lowering of LODs in the following rows: 488 pg/mL (conventional LFIA with Au NPs), 61 pg/mL (silver enhancement), 8 pg/mL (galvanic replacement), and 1 pg/mL (gold enhancement). Using gold enhancement as the optimal technique, the maximal dilution of inactivated SARS-CoV-2-containing samples increased 500 times. The developed LFIA provided highly sensitive and rapid (8 min) point-of-need testing. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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18 pages, 9139 KiB

Open AccessArticle

IQVision: An Image-Based Evaluation Tool for Quantitative Lateral Flow Immunoassay Kits

by Lalitha Pratyusha Bheemavarapu, Malay Ilesh Shah, Jayaraj Joseph and Mohanasankar Sivaprakasam

Biosensors 2021, 11(7), 211; https://doi.org/10.3390/bios11070211 - 28 Jun 2021

Cited by 4 | Viewed by 2972

Abstract

The development of quantitative lateral flow immunoassay test strips involves a lot of research from kit manufacturers’ standpoint. Kit providers need to evaluate multiple parameters, including the location of test regions, sample flow speed, required sample volumes, reaction stability time, etc. A practical [...] Read more.

The development of quantitative lateral flow immunoassay test strips involves a lot of research from kit manufacturers’ standpoint. Kit providers need to evaluate multiple parameters, including the location of test regions, sample flow speed, required sample volumes, reaction stability time, etc. A practical visualization tool assisting manufacturers in this process is very much required for the design of more sensitive and reliable quantitative LFIA test strips. In this paper, we present an image-based quantitative evaluation tool determining the practical functionality of fluorescence-labelled LFIA test cartridges. Image processing-based algorithms developed and presented in this paper provide a practical analysis of sample flow rates, reaction stability times of samples under test, and detect any abnormalities in test strips. Evaluation of the algorithm is done with Glycated Hemoglobin (HbA1C) and Vitamin D test cartridges. Practical sample flow progress for HbA1C test cartridges is demonstrated. The reaction stability time of HbA1C test samples is measured to be 12 min, while that of Vitamin D test samples is 24 min. Experimental evaluation of the abnormality detection algorithm is carried out, and sample flow abnormalities are detected with 100% accuracy while membrane irregularities are detected with 96% accuracy. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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12 pages, 1427 KiB

Open AccessArticle

Polystyrene Microsphere-Based Immunochromatographic Assay for Detection of Aflatoxin B₁ in Maize

by Jin Wang, Xiangmei Li, Xing Shen, Ang Zhang, Jinxiu Liu and Hongtao Lei

Biosensors 2021, 11(6), 200; https://doi.org/10.3390/bios11060200 - 20 Jun 2021

Cited by 9 | Viewed by 2871

Abstract

Aflatoxin B₁ (AFB₁), a mycotoxin, is hepatotoxic, carcinogenic, and nephrotoxic in humans and animals, and contaminate a wide range of maize. In this study, an immunochromatographic assay (ICA) based on polystyrene microspheres (PMs) was developed for sensitive and quantitative detection [...] Read more.

Aflatoxin B₁ (AFB₁), a mycotoxin, is hepatotoxic, carcinogenic, and nephrotoxic in humans and animals, and contaminate a wide range of maize. In this study, an immunochromatographic assay (ICA) based on polystyrene microspheres (PMs) was developed for sensitive and quantitative detection of AFB₁ in maize. The amounts of PMs, the condition for activating carboxyl groups of PMs, the amount of monoclonal antibody (mAb), and the volume of the immune probe were optimized to enhance the performance PMs-ICA for point-of-care testing of AFB₁ in maize. The PMs-ICA showed the cut-off value of 1 ng/mL in phosphate buffer (PB) and 6 µg/kg in maize samples, respectively. The quantitative limit of detection (qLOD) was 0.27 and 1.43 µg/kg in PB and maize samples, respectively. The accuracy and precision of the PMs-ICA were evaluated by analysis of spiked maize samples with recoveries of 96.0% to 107.6% with coefficients of variation below 10%. In addition, the reliability of PMs-ICA was confirmed by the liquid chromatography-tandem mass spectrometry method. The results indicated that the PMs-ICA could be used as a sensitive, simple, rapid point-of-care testing of AFB₁ in maize. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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13 pages, 3349 KiB

Open AccessArticle

Development of a Prototype Lateral Flow Immunoassay of Cortisol in Saliva for Daily Monitoring of Stress

by Elizaveta Panfilova

Biosensors 2021, 11(5), 146; https://doi.org/10.3390/bios11050146 - 07 May 2021

Cited by 8 | Viewed by 3164

Abstract

Emotional stress negatively affects the quality of a person’s daily life. From a physiological point of view, stress is expressed in the excitation of the hypothalamic–pituitary–adrenal cortex axis, which leads to the release of the hormone cortisol into the blood. We developed a [...] Read more.

Emotional stress negatively affects the quality of a person’s daily life. From a physiological point of view, stress is expressed in the excitation of the hypothalamic–pituitary–adrenal cortex axis, which leads to the release of the hormone cortisol into the blood. We developed a lateral flow immunoassay to detect cortisol in human salivary fluid and tested it on 10 healthy volunteers daily for about one month (n = 293 saliva samples). Cortisol was detected in concentrations ranging from 1 to 70 ng/mL. Salivary cortisol levels were confirmed by ELISA. The straightness range of LFIA calibration was from 1 to 100 ng/mL. The diagnostic sensitivity of the method was 73%. It was found that in 3 out of 10 subjects, fluctuations in the level of cortisol in saliva partially corresponded to the subjectively assessed level of stress. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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10 pages, 1898 KiB

Open AccessArticle

Direct Bioelectrocatalytic Oxidation of Glucose by Gluconobacter oxydans Membrane Fractions in PEDOT:PSS/TEG-Modified Biosensors

by Anna Kitova, Sergei Tarasov, Yulia Plekhanova, Aleksandr Bykov and Anatoly Reshetilov

Biosensors 2021, 11(5), 144; https://doi.org/10.3390/bios11050144 - 06 May 2021

Cited by 9 | Viewed by 2322

Abstract

Recent years have witnessed an ever-increasing interest in developing electrochemical biosensors based on direct electron transfer-type bioelectrocatalysis. This work investigates the bioelectrocatalytic oxidation of glucose by membrane fractions of Gluconobacter oxydans cells on screen-printed electrodes modified with thermally expanded graphite and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS). [...] Read more.

Recent years have witnessed an ever-increasing interest in developing electrochemical biosensors based on direct electron transfer-type bioelectrocatalysis. This work investigates the bioelectrocatalytic oxidation of glucose by membrane fractions of Gluconobacter oxydans cells on screen-printed electrodes modified with thermally expanded graphite and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS). Electrooxidation of glucose was shown to occur without the presence of electron transport mediators. Chronoamperometric and cyclic voltametric characteristics showed an increase of anodic currents at electrode potentials of 0–500 mV relative to the reference electrode (Ag/AgCl). The direct electron transfer effect was observed for non-modified PEDOT:PSS as well as for PEDOT:PSS linked with crosslinkers and conductive fillers such as polyethylene glycol diglycidyl or dimethyl sulfoxide. Bioelectrodes with this composite can be successfully used in fast reagent-free glucose biosensors. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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12 pages, 11139 KiB

Open AccessArticle

A Facile Aptasensor for Instantaneous Determination of Cadmium Ions Based on Fluorescence Amplification Effect of MOPS on FAM-Labeled Aptamer

by Yang Liu, Dongwei Zhang, Jina Ding, Kashif Hayat, Xijia Yang, Xuejia Zhan, Dan Zhang, Yitong Lu and Pei Zhou

Biosensors 2021, 11(5), 133; https://doi.org/10.3390/bios11050133 - 23 Apr 2021

Cited by 18 | Viewed by 2558

Abstract

Analytical performance and efficiency are two pivotal issues for developing an on-site and real-time aptasensor for cadmium (Cd²⁺) determination. However, suffering from redundant preparations, fabrications, and incubation, most of them fail to well satisfy the requirements. In this work, we found [...] Read more.

Analytical performance and efficiency are two pivotal issues for developing an on-site and real-time aptasensor for cadmium (Cd²⁺) determination. However, suffering from redundant preparations, fabrications, and incubation, most of them fail to well satisfy the requirements. In this work, we found that fluorescence intensity of 6-carboxyfluorescein(FAM)-labeled aptamer (FAM-aptamer) could be remarkably amplified by 3-(N-morpholino)propane sulfonic acid (MOPS), then fell proportionally as Cd²⁺ concentration introduced. Importantly, the fluorescence variation occurred immediately after addition of Cd²⁺, and would keep stable for at least 60 min. Based on the discovery, a facile and ultra-efficient aptasensor for Cd²⁺ determination was successfully developed. The sensing mechanism was confirmed by fluorescence pattern, circular dichroism (CD) and intermolecular interaction related to pK_a. Under the optimal conditions, Cd²⁺ could be determined rapidly from 5 to 4000 ng mL⁻¹. The detection limit (1.92 ng mL⁻¹) was also lower than the concentration limit for drinking water set by WHO and EPA (3 and 5 ng mL⁻¹, respectively). More than a widely used buffer, MOPS was firstly revealed to have fluorescence amplification effect on FAM-aptamer upon a given context. Despite being sensitive to pH, this simple, high-performance and ultra-efficient aptasensor would be practical for on-site and real-time monitoring of Cd²⁺. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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10 pages, 3086 KiB

Open AccessEditor’s ChoiceArticle

An Electrochemical Aptasensor for Pb²⁺ Detection Based on Metal–Organic-Framework-Derived Hybrid Carbon

by Jina Ding, Dongwei Zhang, Yang Liu, Xuejia Zhan, Yitong Lu, Pei Zhou and Dan Zhang

Biosensors 2021, 11(1), 1; https://doi.org/10.3390/bios11010001 - 22 Dec 2020

Cited by 17 | Viewed by 3607

Abstract

A new double-shelled carbon nanocages material was synthesized and developed an aptasensor for determining Pb²⁺ in aqueous solution. Herein, nanoporous carbon materials derived from core–shell zeolitic imidazolate frameworks (ZIFs) demonstrated excellent electrochemical activity, stability, and high specificity surface area, consequently resulting in [...] Read more.

A new double-shelled carbon nanocages material was synthesized and developed an aptasensor for determining Pb²⁺ in aqueous solution. Herein, nanoporous carbon materials derived from core–shell zeolitic imidazolate frameworks (ZIFs) demonstrated excellent electrochemical activity, stability, and high specificity surface area, consequently resulting in the strong binding with aptamers. The aptamer strands would be induced to form G-quadruplex structure when Pb²⁺ was introduced. Under optimal conditions, the aptasensor exhibited a good linear relationship of Pb²⁺ concentration ranging from 0.1 to 10 μg L⁻¹ with the detection limits of 0.096 μg L⁻¹. The feasibility was proved by detecting Pb²⁺ in spiked water samples and polluted soil digestion solution. The proposed aptasensor showed excellent selectivity and reproducibility, indicating promising applications in environmental monitoring. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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11 pages, 2860 KiB

Open AccessArticle

Label-Free and Sensitive Determination of Cadmium Ions Using a Ti-Modified Co₃O₄-Based Electrochemical Aptasensor

by Yang Liu, Dongwei Zhang, Jina Ding, Kashif Hayat, Xijia Yang, Xuejia Zhan, Dan Zhang, Yitong Lu and Pei Zhou

Biosensors 2020, 10(12), 195; https://doi.org/10.3390/bios10120195 - 30 Nov 2020

Cited by 11 | Viewed by 2330

Abstract

The current work demonstrates an electrochemical aptasensor for sensitive determination of Cd²⁺ based on the Ti-modified Co₃O₄ nanoparticles. In this unlabeled system, Ti-modified Co₃O₄ nanoparticles act as current signal amplifiers modified on the screen-printed carbon electrode [...] Read more.

The current work demonstrates an electrochemical aptasensor for sensitive determination of Cd²⁺ based on the Ti-modified Co₃O₄ nanoparticles. In this unlabeled system, Ti-modified Co₃O₄ nanoparticles act as current signal amplifiers modified on the screen-printed carbon electrode (SPCE) surface, while the derivative aptamer of Cd²⁺ works as a target recognizer. In addition, the sensing is based on the increase in electrochemical probe thionine current signal due to the binding of aptamer to Cd²⁺ via specific recognition. In the current study, key parameters, including aptamer concentration, pH, and incubation time were optimized, respectively, to ensure sensing performance. Cyclic voltammetry was used not only to characterize each preparation and optimization step, but also to profile the bindings of aptamer to Cd²⁺. Under optimal conditions, Cd²⁺ can be determined in a linear range of 0.20 to 15 ng/mL, with a detection limit of 0.49 ng/mL, significantly below the maximum concentration limit set by the U.S. Environmental Protection Agency. Based on comparative analysis and the results of recovery test with real samples, this simple, label-free but highly selective method has considerable potential and thus can be used as an in-situ environmental monitoring platform for Cd²⁺ testing. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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12 pages, 723 KiB

Open AccessPerspective

Glycated Hemoglobin and Methods for Its Point of Care Testing

by Miroslav Pohanka

Biosensors 2021, 11(3), 70; https://doi.org/10.3390/bios11030070 - 04 Mar 2021

Cited by 21 | Viewed by 7362

Abstract

Glycated hemoglobin (HbA_1c) is a product of the spontaneous reaction between hemoglobin and elevated glucose levels in the blood. It is included among the so-called advanced glycation end products, of which is the most important for the clinical diagnosis of diabetes [...] Read more.

Glycated hemoglobin (HbA_1c) is a product of the spontaneous reaction between hemoglobin and elevated glucose levels in the blood. It is included among the so-called advanced glycation end products, of which is the most important for the clinical diagnosis of diabetes mellitus, and it can serve as an alternative to glycemia measurement. Compared to the diagnosis of diabetes mellitus by glycemia, the HbA_1c level is less influenced by a short-term problem with diabetes compensation. Mass spectroscopy and chromatographic techniques are among the standard methods of HbA_1c level measurement. Compared to glycemia measurement, there is lack of simple methods for diabetes mellitus diagnosis by means of the HbA_1c assay using a point-of-care test. This review article is focused on the surveying of facts about HbA_1c and its importance in diabetes mellitus diagnosis, and surveying standard methods and new methods suitable for the HbA_1c assay under point-of-care conditions. Various bioassays and biosensors are mentioned and their specifications are discussed. Full article

(This article belongs to the Special Issue New Developments for Efficient Rapid Bioassays)

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