*4.2. Inventory of Commercially Available Kits for Mycotoxins Detection*

Commercial test kits for mycotoxin detection are utilized as an appropriate alternative for more user-friendly, inexpensive, robust, and rapid analysis. There are a large number of commercial detection kits available for mycotoxin analysis in the current market, as summarized in the Table 6. They commonly include ELISA kits, membrane-based immunoassays such as lateral flow immunoassays (LFIAs), fluorescence polarization immunoassays (FPIAs), and immunoaffinity column coupled with fluorometric assay. The majority of these test kits are based on an immunoassay format which relies on the specific interaction of antigen and antibody. Moreover, colorimetric detection kits are most preferred because of the ability to see results with the naked eye. Among them, LFIAs

with acceptable sensitivity, good accuracy, portability, short detection time, ease of use, and no need for specialized personnel have become strong competitors on the market for mycotoxin analysis. Several companies worldwide produce LFIA test strip to analyze different kinds of mycotoxins. Charm Sciences Inc., Pribolab, EnviroLogix, Romer labs, Vicam, CUSABIO, etc. are among LFIA strips producer for aflatoxins, DON, ZEN, T2, and OTA. ROSA (rapid one step assay) lateral flow strips developed by Charm Sciences Inc. are the leading mycotoxin test worldwide.

Commercial LFIAs test strips usually use AuNPs as colored label. The method can provide qualitative and/or semi-quantitative results within minutes (e.g., Afla-V and AflaCheck strip tests by Vicam, 5 and 3 min, respectively). For semi-quantitative analysis, portable readers have been developed for on-site detection. For example, PerkinElmer's QuickSTAR Horizon strip reader provides quantitative results for mycotoxins including AFB1, AFB2, AFG1, and AFG2; at detection levels of 2 to 300 ppb within 6 min. Charm EZ-M Reader is another type of portable strip reader which can show results within 3–5 min. The color-coded strips allow the EZ-M reader to automatically recognize which mycotoxin group you are testing and will adjust the reader temperature and time accordingly. Some companies such as R-Biopharm provided a mobile app on a smartphone to analyze color signal instead of a reader, specifically for aflatoxins, T2/HT2, ZEN, and FMN.

Commercially available test kits have been developed for determination of individual mycotoxins or for multiple mycotoxins in one group (e.g., aflatoxins B1, B2, G1, and G2). Since we are usually faced with contamination of food and feed to more than one mycotoxin, the current trend in LFIA technology is to develop strips with multiple test lines for the simultaneous detection of multiple mycotoxins.

After LFIA test strips, ELISA-based kits have allocated a major portion of market amongst other mycotoxin detection methods. Many companies—such as Sigma, Elabscience, Eurofins, Romer Labs, ELISA Technologies, Cusabio, Astori Tecnica, etc.—offer ELISA kits to detect the most common types of mycotoxins. Some of these kits can detect several types of mycotoxins in one group (e.g., aflatoxins B1, B2, G1, and G2).




The HRP-TMB system is the most common method for colorimetric signal generation in ELISA-based kits. Commercial ELISA kits are sensitive, selective, high throughput, with minimum sample preparation steps. Moreover, the detection time in commercial kits has been shortened so that, most of them are able to detect target mycotoxin within 1–2 h. Romer labs has produced ELISA kits for aflatoxins, OTA, DON, T2, ZEN, and FMN with incubation periods of 15 min. In some kits, cross-reactivity of antibodies leads to overestimation of results while matrix effect plays a key role in providing false-positive results. To avoid such effects, most kits define the limited matrices to which the ELISA kit can be used [115]. However, this feature can be considered a limitation for such ELISA kits. On the other hand, some companies such as Eurofins have developed sensitive ELISA method for a wide range of matrices.

As with the LFIA kits, the current trend in ELISA technique is to develop commercial multiplex assays. Multiplex ELISA can be developed by immobilizing different toxins in different wells of a single microplate in competitive format.
