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      Fabrication of microfluidic device for Aflatoxin M1 detection in milk samples with specific aptamers

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          Abstract

          This study describes the colorimetric detection of aflatoxin M1 (Afl M1) in milk samples using a microfluidic paper-based analytical device (µPAD). Fabrication of µPADs was done using a simple and quick approach. Each μPAD contained a detection zone and a sample zone interconnected by microchannels. The colorimetric assay was developed using unmodified AuNPs as a probe and 21-mer aptamer as a recognition molecule. The free aptamers were adsorbed onto the surface of AuNPs in absence of Afl M1, even at high salt concentrations. The salt induced aggregation of specific aptamers occurred in presence of Afl M1. Under optimum conditions, the analytical linear range was found to be 1 µM to 1 pM with limit of detection 3 pM and 10 nM in standard buffer and spiked milk samples respectively. The proposed aptamer based colorimetric assay was repeatable, quick, selective, and can be used for on-site detection of other toxins in milk and meat samples.

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          Aptamers and Their Biological Applications

          Recently, aptamers have attracted the attention of many scientists, because they not only have all of the advantages of antibodies, but also have unique merits, such as thermal stability, low cost, and unlimited applications. In this review, we present the reasons why aptamers are known as alternatives to antibodies. Furthermore, several types of in vitro selection processes, including nitrocellulose membrane filtration, affinity chromatography, magnetic bead, and capillary electrophoresis-based selection methods, are explained in detail. We also introduce various applications of aptamers for the diagnosis of diseases and detection of small molecules. Numerous analytical techniques, such as electrochemical, colorimetric, optical, and mass-sensitive methods, can be utilized to detect targets, due to convenient modifications and the stability of aptamers. Finally, several medical and analytical applications of aptamers are presented. In summary, aptamers are promising materials for diverse areas, not just as alternatives to antibodies, but as the core components of medical and analytical equipment.
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            Mycotoxins and their effects on human and animal health

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              Ultrasensitive electrochemical immuno-sensing platform based on gold nanoparticles triggering chlorpyrifos detection in fruits and vegetables.

              Chlorpyrifos (chl) is an organophosphate pesticide extensively used in agriculture and highly toxic for human health. Fluorine doped tin-oxide (FTO) based electrochemical nanosensor was developed for chlorpyrifos detection with gold nanoparticles (AuNPs) and anti-chlorpyrifos antibodies (chl-Ab). AuNPs provides high electrical conductivity and specific resistivity, thus increases the sensitivity of immunoassay. High electrical conductivity of AuNPs reveals that it promotes the redox reaction for better cyclic voltammetry. Based on the intrinsic conductive properties of FTO-AuNPs complex, chl-Ab was immobilized onto AuNPs surface. Under optimized conditions, the proposed FTO based nanosensor exhibited high sensitivity and stable response for the detection of chlorpyrifos, ranging from 1fM to 1µM with limit of detection (LOD) up to 10fM. The FTO-AuNPs sensor was successfully employed for the detection of chlorpyrifos in standard as well in real samples up to 10nM for apple and cabbage, 50nM for pomegranate. The proposed FTO-AuNPs nanosensor can be used as a quantitative tool for rapid, on-site detection of chlorpyrifos traces in real samples when miniaturized due to its excellent stability, sensitivity, and simplicity.
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                Author and article information

                Contributors
                sonugandhi@gmail.com
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                13 March 2020
                13 March 2020
                2020
                : 10
                : 4627
                Affiliations
                [1 ]DBT- National Institute of Animal Biotechnology, Hyderabad, 500032 India
                [2 ]Department of Biotechnology, JNTUA College of Engineering, Andhra Pradesh, 516390 India
                [3 ]ISNI 0000 0004 1773 5396, GRID grid.56302.32, Department of Pharmaceutical Chemistry, , King Saud University, ; Riyadh, 11451 Saudi Arabia
                [4 ]Department of Biomedical Engineering, Indian Institute of Technology (IIT), Hyderabad, 502285 India
                Article
                60926
                10.1038/s41598-020-60926-2
                7070014
                32170077
                d1b02db1-3056-4154-92c5-746f73a7ff7f
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 7 January 2020
                : 5 February 2020
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                © The Author(s) 2020

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                biological techniques,nanoscience and technology
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                biological techniques, nanoscience and technology

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