Multimycotoxin dipstick enzyme immunoassay applied to wheat

Abstract

A membrane-based visual dipstick enzyme immunoassay for the simultaneous detection of up to five mycotoxins was developed. Multiple dots of the respective antibodies against aflatoxin B1 (AFB1), T-2 toxin (T-2), 3-acetyldeoxynivalenol (3-AcDON), roridin A (RA), and zearalenone (ZEA) were applied onto a dipstick membrane. The competitive immunoreactions were performed by incubation of the dipstick in a test tube containing sample solution and a mixture of the respective toxin-horseradish peroxidase conjugates. After a second incubation in enzyme substrate/chromogen solution, a complete suppression of the (blue) colour development of the respective dot was scored positive. Visual detection limits for AFB1, T-2, 3-AcDON, RA, and ZEA in buffer solution were 2 ng/ml, 5 ng/ml, 50 ng/ml, 30 ng/ml, and 5 ng/ml respectively. Using a simple extraction procedure, the detection limits of the multimycotoxin assay in artificially contaminated wheat samples were 30 ng/g, 100 ng/g, 600 ng/g, 500 ng/g, and 60 ng/g, respectively.

Advances in the analysis of mycotoxins and its quality assurance

“…Qualitative and semi-quantitative results can also be obtained by lesser experienced personnel. Multi-analyte dipstick immunoassays for various mycotoxins have been developed, however, with limited sensitivity (Schneider et al 1995). Another interesting alternative rapid assay format for the detection of mycotoxins are lateral flow devices.…”

Surface Plasmon Resonance‐Based Immunoassay for the Detection of Aflatoxin B1Using Single‐Chain Antibody Fragments

“…Daly and collaborators developed a Biacore-based assay using a polyclonal antibody in PBS with a limit of detection of 3 ng mL 21 [10] and an scFv-based Biacore assay with a limit of detection of 3 ng mL 21 in PBS and 0.75 ng mL 21 in spiked grain. [17] The limits of detection described in this paper also compare favorably with several other immunoassay formats including a fluorescence polarization assay for aflatoxins with a range of detection between 5 and 200 ppb, [27] a dipstick assay with a limit of detection of 2 ng mL 21 , [28] and ELISA formats described by Candlish et al, [29] Aldao et al, [30] and Daly et al [10,17] with limits of detection at 0.2 ng mL 21 , 0.25 mg kg 21 , 3 ng mL 21 , and 98 ng mL 21 , respectively. Analytical techniques, including the HPLC detection system described by Kussak et al [31] have offered greater sensitivity with limits of detection for aflatoxins B 1 , B 2 , G 1 , and G 2 at 6.8 pg mL 21 in urine, a sol-particle lateral flow immunoassay with limits of detection of 0.1 ppb in buffer and 10 ppb in grain samples, [32] and an immunoaffinity fluorometric biosensor with a lower limit of detection, at 0.1 ppb.…”

Bioaerosol lung damage in a worker with repeated exposure to fungi in a water-damaged building.

“…The standard used to associate health effects to mycotoxin exposure is the measurement of mycotoxins in bulk samples obtained from materials such as walls, ceiling tiles, and air ducts (12,13). Although the identification and quantification of several mycotoxins has been recently accomplished using monoclonal antibodies and enzyme immunoassays, these methods have had limited use in field evaluations (10,(14)(15)(16). The pilot serologic survey of this investigation was not able to distinguish between exposed and unexposed workers in a building heavily contaminated with mycotoxin-producing fungi.…”