T-2 Toxin/HT-2 Toxin and Ochratoxin A ELISAs Development and In-House Validation in Food in Accordance with Commission Regulation (EU) No 519/2014

Abstract

T-2 toxin/HT-2 toxin (T-2/HT-2) and ochratoxin A (OTA) are mycotoxins that can contaminate a variety of agricultural commodities. To protect consumers’ health, indicative limits for T-2/HT-2 and maximum limits for OTA have been set by the European Commission, requiring food business operators and controlling agencies to conduct routine checks for the presence of these harmful contaminants. Screening methods are increasingly used for monitoring purposes. Due to the demand for new and improved screening tools, two individual detection methods, T-2/HT-2 and OTA enzyme-linked immunosorbent assays (ELISAs), were developed in this study. The T-2/HT-2 ELISA was based on a T-2 monoclonal antibody with an IC50 (50% inhibitory concentration) of 0.28 ng/mL and 125% cross-reactivity with HT-2. As regards the OTA ELISA, a new sensitive monoclonal antibody specific to OTA with an IC50 of 0.13 ng/mL was produced. Both developed ELISA tests were then validated in agricultural commodities in accordance with the new performance criteria guidelines for the validation of screening methods for mycotoxins included in Commission Regulation (EU) No 519/2014. The T-2/HT-2 ELISA was demonstrated to be suitable for the detection of T-2/HT-2 in cereals and baby food at and above the screening target concentration (STC) of 12.5 μg/kg and 7.5 μg/kg, respectively. The OTA ELISA was shown to be applicable for the detection of OTA in cereals, coffee, cocoa and wine at and above the STC of 2 μg/kg, 2.5 μg/kg, 2.5 μg/kg and 0.4 ng/mL, respectively. The accuracy of both ELISAs was further confirmed by analysing proficiency test and reference samples. The developed methods can be used for sensitive and high-throughput screening for the presence of T-2/HT-2 and OTA in agricultural commodities.

ELISA and UPLC/FLD as Screening and Confirmatory Techniques for T-2/HT-2 Mycotoxin Determination in Cereals

“…657/2002, this parameter has to be lower than 23% and 16% for the fortification levels equal to 100 and 250 μg kg −1 , respectively. This parameter was comparable with the precision of ELISA obtained by Pleadin et al [21], who analyzed the same types of cereals samples investigated in the present study, and higher that that reported by Oplatowska-Stachowiak et al [22] for rye and baby porridge analysis. The recovery percentage was satisfactory as well, since both fortification levels gave a mean value of 102%, whereas the required range set in Decision No.…”

Layered material platform for surface plasmon resonance biosensing

“…Both toxins are produced by moulds that grows on improperly stored grains 12 . HT-2 can cause acute or chronic intoxication of humans and animals 39,40 . The ability to detect low quantities of HT-2 is therefore of great interest for food safety and it can be performed by conventional SPR techniques based on Au with limited sensitivity (~ng/mL) as reported in ref.…”

Quantum dots‐based fluorescence immunoassay for detection of tiamulin in pork

“…The inhibitory concentration was defined as (1−B/B0) × 100%. Then the half inhibitory concentration (IC 50 , defined as 50% inhibitory concentration), limit of detection (LOD, IC 10 , defined as 10% inhibitory concentration) and detection range (defined as ranged from IC 20 to IC 80 ) were calculated according to the standard curve (Oplatowska‐Stachowiak et al, 2017).…”

“…In addition, these techniques are hindered by low sample throughput and long analysis time. The immunological methods for TML detection have been used widely in recent years due to their advantages of simple, rapid, and on‐site detection (Deng et al, 2017; Liu, Chu, & Yu, 2016; Oplatowska‐Stachowiak et al, 2017).…”