DESARROLLO DE UN SISTEMA DE DETECCIÓN DE OCRATOXINA A BASADO EN NANOSENSORES APTAMÉRICOS EN MICROPLACAS

Angeline Saldaña; Yulán Hernandez; Betty Galarreta

Autores/as

Angeline Saldaña Pontificia Universidad Católica del Perú, Departamento de Ciencias-Sección Química, Lima, Perú https://orcid.org/0000-0003-4105-071X
Yulán Hernandez Pontificia Universidad Católica del Perú, Departamento de Ciencias-Sección Química, Lima, Perú https://orcid.org/0000-0003-1276-5927
Betty Galarreta Pontificia Universidad Católica del Perú, Departamento de Ciencias-Sección Química, Lima, Perú https://orcid.org/0000-0003-4860-3161

Resumen

La ocratoxina A (OTA) es una micotoxina altamente tóxica producida por algunas variedades de hongos, capaz de contaminar productos agroindustriales, lo que ha obligado a establecer normas de regulación en varios países. Por tal motivo, es necesario contar con métodos confiables y sensibles para su detección temprana, que puedan ser utilizados de manera rutinaria. Los nanosensores aptaméricos representan una alternativa que puede mejorar la detección de moléculas de interés, debido a la alta afinidad y selectividad de los biorreceptores aptaméricos por objetivos específicos. Además, el uso de nanomateriales de metales nobles permite desarrollar sensores a partir de sus propiedades ópticas, como la resonancia de plasmón superficial localizado (LSPR), al poder monitorear los cambios en la superficie y relacionarlo con la concentración de una sustancia de interés. En este trabajo se muestran los avances en el desarrollo de un sistema alternativo de detección de OTA basado en nanosensores aptaméricos. El procedimiento contempla la síntesis y funcionalización de nanoesferas de oro (AuNEs) con un aptámero selectivo a OTA (APT), el proceso de modificación de microplacas con la hebra complementaria a dicho aptámero (HC) y la hibridación entre los oligonucleótidos conjugados. Finalmente, a modo de prueba de concepto, se realizaron algunas pruebas que permitieron discriminar la presencia de 336 ppb de OTA.

Palabras clave: Aptámeros, micotoxinas, microplacas, nanosensores, ocratoxina A, oro.

Abstract

Ochratoxin A (OTA) is a highly toxic mycotoxin produced by some varieties of fungi, capable of contaminating agro-industrial products, this has forced the establishment of regulatory standards in several countries. Therefore, it is required to have reliable and sensitive methods for its early detection that could be implemented in routine assays. Aptameric nanosensors are an alternative method that can improve the detection of specific target molecules, due to the high affinity and selectivity of aptameric bioreceptors for specific targets. In addition, the use of noble metal nanomaterials allows the development of sensors based on their optical properties such as localized surface plasmon (LSPR), as it is possible to monitor changes in their surface and relate them to the concentration of a target analyte. This work has focused in the development of an alternative OTA detection system, based on aptameric nanosensors. The procedure contemplates the synthesis and functionalization of gold nanospheres (AuNEs) with an OTA-selective aptamer (APT), a microplate surface modification procedure with the complementary strand to the aptamer (HC) and the hybridization between the oligonucleotides. Lastly, as a proof of concept, some assays were performed to discriminate the presence of 336 ppb of OTA.

Keywords: Aptamers, gold, microplates, mycotoxins, nanosensors, ochratoxin A.

*Autor para correspondencia: E. mail: bgalarreta@pucp.pe

DOI: http://dx.doi.org/10.17268/rebiol.2021.41.01.03

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