Development of an Immunofluorescence Assay Module for Determination of the Mycotoxin Zearalenone in Water

Abstract

Project Aquafluosense is designed to develop prototypes for a fluorescence-based instrumentation setup for in situ measurements of several characteristic parameters of water quality. In the scope of the project an enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the detection of several environmental xenobiotics, including mycotoxin zearalenone (ZON). ZON, produced by several plant pathogenic Fusarium species, has recently been identified as an emerging pollutant in surface water, presenting a hazard to aquatic ecosystems. Due to its physico-chemical properties, detection of ZON at low concentrations in surface water is a challenging task. The 96-well microplate-based fluorescence instrument is capable of detecting ZON in the concentration range of 0.09–400 ng/mL. The sensitivity and accuracy of the analytical method has been demonstrated by a comparative assessment with detection by high-performance liquid chromatography and by total internal reflection ellipsometry. The limit of detection of the method, 0.09 ng/mL, falls in the low range compared to the other reported immunoassays, but the main advantage of this ELFIA method is its efficacy in combined in situ applications for determination of various important water quality parameters detectable by induced fluorimerty—e.g., total organic carbon content, algal density or the level of other organic micropollutants detectable by immunofluorimetry. In addition, the immunofluorescence module can readily be expanded to other target analytes if proper antibodies are available for detection.

In vitro and in vivo evaluation of AFB1 and OTA-toxicity through immunofluorescence and flow cytometry techniques: A systematic review

“…Furthermore, FCM provides an excellent interpretation of cellular processes such as redox balance, proliferation and apoptosis (Zhou et al, 2020;Broemsen et al, 2021). Whilst, microscopy, which is a rapid, cheap and simple technic, allows to investigate and examine histological changes on tissues induced by mycotoxins and through specific antibodies and fluorescence kits it enables to analyze their specific MOA (Nguyen et al, 2019;Gémes et al, 2021).…”

Utilization of a Novel Immunofluorescence Instrument Prototype for the Determination of the Herbicide Glyphosate

“…The scheme of this immunofluorescence module is shown in Figure 4 . The motor, optics and sample holder were developed at Optimal Optik Ltd. (Budapest, Hungary), while the detector electronics were created at the Budapest University of Technology and Economics (Budapest, Hungary) and described by Gémes et al [ 34 ]. Briefly, the samples were illuminated by an LED light source with a 520 to 535 nm minimum to maximum dominant wavelength range (Cree XPEBGR-L1-0000-00F01; Cree LED, Durham, NC, USA), and the emission was detected by a dichroic beam path with silicon photodiodes (PIN-25D, OSI Optoelectronics; Hawthorne, CA, USA).…”

Mycotoxins as Emerging Contaminants. Introduction to the Special Issue “Rapid Detection of Mycotoxin Contamination”

“…The LOD of the method is 1.1 ng/mL, comparable to a laboratory-based enzyme-linked immunosorbent assay (ELISA) method with a LOD of 0.29–0.39 ng/mL. The development of a portable instrument for ZEN by enzyme-linked fluorescent immunoassay (ELFIA) is reported by Gémes et al [ 15 ], but as this instrument is a novel application for detection of mycotoxins as emerging water contaminants, it is discussed among the applications of routine monitoring (see Section 3 . Applications in routine monitoring).…”

“…The occasional occurrence of mycotoxins in surface and drinking water is not a newly identified phenomenon, but its particular significance has been emphasized lately [ 7 , 8 , 22 , 24 , 25 , 26 ], classifying mycotoxins and their metabolites as emerging surface water contaminants [ 7 , 27 ], and assessing their routes of occurrence [ 28 , 29 ]. Gémes et al [ 15 ] report the development of an ELFIA method as a module of a portable, in situ fluorimeter instrument installed in a mobile laboratory vehicle to detect ZEN in water with a LOD of 0.09 ng/mL. This LOD appears to be quite favorable compared to reported ELISAs, but a major advantage of the ELFIA method lies in its combined in situ applicability in the determination of important water quality parameters detectable by induced fluorimetry—e.g., total organic carbon content, algal density or the level of other organic micropollutants.…”