A flow cytometry based competitive fluorescent microsphere immunoassay (CFIA) system for detecting up to six mycotoxins

Czéh, Árpád; Mandy, Francis; Feher-Toth, Szilvia; Torok, Livia; Mike, Zoltan; Kőszegi, Balázs; Lustyik, György

doi:10.1016/j.jim.2012.07.010

Cited by 37 publications

(47 citation statements)

References 25 publications

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“…Although the triplex assay principle, described in this work, seems similar to the work of Czeh et al [34], there are some crucial differences. Besides sensitivity (discussed above), we used certified reference materials designated to single mycotoxins (and in case of FB 1 also the FB 2 and FB 3 metabolites) in combination with a confirmatory LC-MS/MS method to get full information about all relevant metabolites present for a comprehensive comparison of the triplex assay results.…”

Section: Resultsmentioning

confidence: 82%

“…For OTA, the sensitivities were 0.7 and 3.4 μg/kg, respectively (ML = 5 μg/kg); for ZEN, 5.8 and 32 μg/kg, respectively (ML = 100 μg/kg); and for FB 1 + FB 2 , 170 and 1270 μg/kg, respectively (ML = 2000 μg/kg). Maize, the material with the highest matrix effect on the dose–response curves, was not tested (or presented) in the work of Czeh et al [34]; therefore, we compared our wheat sensitivities with their results. Our OTA assay was two times more sensitive, while their ZEN and FB 1 assays were respectively four and five times more sensitive.…”

Section: Resultsmentioning

confidence: 99%

“…In contrast, direct immunoassays having immobilised antibodies on the beads are rare. Very recently, Czeh et al [34] described a similar approach as in the present work, but using a different analyser with non-paramagnetic beads. Unfortunately, experimental data as well as detailed procedures are lacking in [34], thus hampering a direct comparison with our results.…”

Section: Introductionmentioning

confidence: 95%

“…Very recently, Czeh et al [34] described a similar approach as in the present work, but using a different analyser with non-paramagnetic beads. Unfortunately, experimental data as well as detailed procedures are lacking in [34], thus hampering a direct comparison with our results. Moreover, no LC-MS/MS or other confirmatory method was carried out using certified reference samples to verify or pre-validate that assay.…”

Section: Introductionmentioning

confidence: 95%

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Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

et al. 2013

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A combined (triplex) immunoassay for the simultaneous detection of three mycotoxins in grains was developed with superparamagnetic colour-encoded microbeads, in combination with two bead-dedicated flow cytometers. Monoclonal antibodies were coupled to the beads, and the amounts of bound mycotoxins were inversely related to the amounts of bound fluorescent labelled mycotoxins (inhibition immunoassay format). The selected monoclonal antibodies were tested for their target mycotoxins and for cross-reactivity with relevant metabolites and masked mycotoxins. In the triplex format, low levels of cross-interactions between the assays occurred at irrelevant high levels only. All three assays were influenced by the sample matrix of cereal extracts to some extent, and matrix-matched calibrations are recommended for quantitative screening purposes. In a preliminary in-house validation, the triplex assay was found to be reproducible, sensitive and sufficiently accurate for the quantitative screening at ML level. The triplex assay was critically compared to liquid chromatography–tandem mass spectrometry using reference materials and fortified blank material. Results for the quantification of ochratoxin A and zearalenone were in good agreement. However, the fumonisin assay was, due to overestimation, only suitable for qualitative judgements. Both flow cytometer platforms (Luminex 100 and FLEXMAP 3D) performed similar with respect to sensitivity with the advantages of a higher sample throughput and response range of the FLEXMAP 3D and lower cost of the Luminex 100.The priciple of the direct inhibition microbead immunoassay using fluorescent mycotoxin-reporter conjugatesElectronic supplementary materialThe online version of this article (doi:10.1007/s00216-013-7095-7) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 95%

See 2 more Smart Citations

Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

et al. 2013

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“…The matrix effects caused by co-elution of matrix components should be carefully considered because they can result in suppression or, more rarely, enhancement of the ionization efficiency of the analyte affecting the accuracy and precision of the results, especial for injection of crude extracts [33,68]. Matrix-matched standards calibration has accurately quantified multiplex mycotoxins in different food matrix without need to use isotope-labelled internal standards [33].…”

Section: Hplc-ms Hplc-ms/ms and Ultra Hplc-ms/ms (Uplc-ms/ms)mentioning

confidence: 99%

High Throughput Detection Methods for Multiplex Mycotoxins

Li¹,

Deng²,

Liu³

et al. 2017

Toxicol Open Access

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Mycotoxins are toxic low-molecular-weight compounds which produced by the metabolism of certain fungi species. Due to their toxicity, chemistry stability, diversity and co-occurrence in agriculture products, it is urgently needed to develop the rapid, simple and effective detection technique methods to monitor and prevent mycotoxin contamination in whole food chain. This article gives a review about high throughput detection methods for multiplex mycotoxins which mainly includes chromatographic instrument techniques, microarray chip, suspension array, lateral flow biosensors, surface plasmon resonance (SPR) and nanoparticle-based biosensors. The advantages and disadvantages and the key steps of them have been discussed. The insight and evaluation of the technique progress were given, which would be helpful to further develop this filed.

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Immunoassay

Taylor¹

2016

Kirk-Othmer Encyclopedia of Chemical Technology

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Immunoassay, a diagnostic and measurement method that utilizes antibodies to detect and quantify specific analytes (antigens), is characterized by high analyte specificity, high sensitivity (routinely detecting ng to pg analyte levels), and rapid data output. It is amenable to a variety of formats, from single immunoassays carried out manually, to fully automated instruments capable of performing multiple assays on the same or different analytes. Immunoassay quantification methods are based on immunoprecipitation, such as agglutination assays, or on specific indicator reagents linked to antibody or antigen reagents used in the immunoassay. These include radioisotopes for radioimmunoassay, enzymes for enzyme‐linked immunoassay, fluorophores for fluorescence immunoassay, and chemicals or proteins for chemiluminescent or bioluminescent immunoassay. Immunoassay technology is continuing to evolve and applications have moved beyond traditional clinical settings to industry and the environment. Immunoassays are also serving as the basis for newer measurement technologies including hybrid immunoassay‐nucleic acid probe assays, immunobiosensors, and antibody‐based microarrays.

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A flow cytometry based competitive fluorescent microsphere immunoassay (CFIA) system for detecting up to six mycotoxins

Cited by 37 publications

References 25 publications

Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

High Throughput Detection Methods for Multiplex Mycotoxins

Immunoassay

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