During shaking, mycotoxins and biotinylated mycotoxin conjugates would compete for binding with their related antibodies. limit of detection (LOD) of this antibody microarray for aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1in corn was 5.25, 4.75, 2.25, and 6 g/kg, respectively. The recovery rates from your spiked samples were between 79.2% and 113.4%, with coefficient of variation <10%. The results of the analysis of commercial samples for mycotoxins by using this fresh assay and the liquid chromatography-tandem mass spectrometry (LC-MS/MS) were similar and in good agreement. This assay could also be revised for the simultaneous detection of additional multiple mycotoxins, as well as low-weight analytes, dangerous to human health. Keywords:mycotoxins, biotin-streptavidin, microarray, quantification == 1. Intro == Mycotoxins, the secondary metabolites produced by fungi such asAspergillus,Fusarium, andPenicillium, often contaminate agricultural create during harvest, storage, or processing [1]. The most frequently experienced and analyzed mycotoxins in cereal grains and feeds are aflatoxins, ochratoxins, zearalenone, and fumonisins [2]. The major toxicities of these mycotoxins include carcinogenicity (aflatoxin B1, AFB1), potentially LIG4 carcinogenic and nephrotoxic providers (ochratoxin A, OTA), estrogenic and reproductive toxicity (zearalenone, ZEN), and equine leucoencephalomalacia and porcine pulmonary edema (fumonisin B1, FB1) [3]. Their common distribution has become a major concern in food and feed security, and simultaneous event of multiple mycotoxins is definitely a common trend. Synergism in toxicities is definitely observed when a multiplex of mycotoxins happens in one product [4,5]. Considering the severe health risk of mycotoxins, many countries and regulatory government bodies, such as the Joint Food and Agriculture Corporation (FAO)/World Health Corporation BML-284 (Wnt agonist 1) (WHO) Expert Committee on Food Additives (JECFA), have defined the maximum residue limit (MRL) in food products in the ppb level (parts per billion or g/kg) [6]. Series of analytical methods have been developed and are in use to quantify the levels of these mycotoxins. Instrumental methods, such as high-performance liquid chromatography (HPLC) [7], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [8], and gas chromatography-mass spectrometry (GC-MS) [9] are highly sensitive and create reliable results. However, these require complex and tedious sample preparatory methods, and are expensive while also becoming time-consuming. Immunoassays, such as multiplex circulation cytometric immunoassay [10], microarray-based methods [11], and fluorescence polarization immunoassay [12], have proven to be excellent methods for multi-component detection because of the advantage of high throughput. The need for special tools and skilled specialists in the detection process restricts their considerable use. Traditional immunoassay, the enzyme-linked immunosorbent assay (ELISA), has been widely used like a screening method due to its high level of sensitivity, rate, and low-cost, but is definitely probably hard to implement like a high-throughput detection method. Promising methods, such as series electrochemical assays, have been employed for the detection of mycotoxins, but significant improvements still need to be made on some important parts (e.g., simplicity, ease of operation, and multi-mycotoxin acknowledgement) to render them an ideal analytical platform in the near future [13]. The need for methods BML-284 (Wnt agonist 1) able to detect multiple-mycotoxin in one assay is the background on which this study is based. Since the development of arrays, this technology has been widely applied in many areas, such as clinical medicine, cell biology, proteomics, and analytical chemistry [14]. Several suspension arrays that use an indirect competition strategy have been reported to be used for detection of multiple mycotoxins [15,16,17]. These methods use lengthy, complex methods, and reactions are carried out in Eppendorf tubes. Solid-phase array is definitely a combination of immunoassay and protein microarrays. In this BML-284 (Wnt agonist 1) system, transmission responses produced by antigen (Ag)-antibody (Ab) connection is observed by a confocal laser scanner and image extraction software. This high-throughput detection platform has the.