Detection of T-2 toxin in different cereals by flow-through enzyme immunoassay with a simultaneous internal reference.
In most previously described membrane-based immunoassays a separate negative control assay is always carried out to evaluate the performance of the assay. To overcome this problem, a membrane-based flow-through enzyme immunoassay with an internal control has been developed for the detection of T-2 toxin in cereals (patent pending). An Immunodyne ABC membrane was coated with 2 microL of goat anti-horseradish peroxidase (HRP) (internal control spot) (1:1000) and 2 microL of rabbit anti-mouse (test spot) (undiluted) immunoglobulins, and the free binding sites were blocked. In addition to the antibody-coated Immunodyne ABC membrane, the assay also comprises a plastic snap-fit device, absorbent cotton wool, mouse anti-T-2 monoclonal antibodies (Mab), and T-2-HRP conjugate. The color intensity (Delta) of the internal control and that of the negative sample showed no difference (P > 0.05), whereas there was a significant difference between the internal control and positive samples (P < 0.05). The minimum detectable limit that could be visually detected with confidence was 50 ng of T-2 per gram of cereal sample.
Sibanda
L
L
Laboratory of Food Analysis, Faculty of Pharmaceutical Sciences, Gent University, Harelbekestraat 72, B-9000 Gent, Belgium. Liberty.Sibanda@rug.ac.be
De Saeger
S
S
Van Peteghem
C
C
Grabarkiewicz-Szczesna
J
J
Tomczak
M
M
D002523:Edible Grain / Q000737:chemistry* / Q000382:microbiology; D005506:Food Contamination; D005670:Fusarium / Q000378:metabolism; D007124:Immunoenzyme Techniques / Q000379:methods*; D008567:Membranes, Artificial; D013605:T-2 Toxin / Q000032:analysis*
D008567:Membranes, Artificial; D013605:T-2 Toxin