Development of a one step strip test for the detection of sulfadimidine residues†
The one step strip test described is a competitive immunoassay in which the detector reagent consists of colloidal gold particles coated with affinity purified polyclonal anti-sulfadimidine (SDD) antibodies. The capture reagent in the assay is an SDD–ovalbumin conjugate which is immobilised on the lateral flow membrane of the test device. In the test procedure, 150 µl (four drops) of a liquid sample (buffer, urine or milk) are brought into the sample well of the test device and allowed to migrate over the membrane. The more analyte present in the sample, the more effectively it will compete with the SDD immobilised on the membrane for binding to the limited amount of antibodies of the detector reagent. A sufficient amount of SDD in the sample will therefore prevent the binding of the detector reagent to the SDD immobilised on the membrane. Therefore, a positive sample will not show a test line in the read-out zone. With spiked buffer or calf urine this was obtained at a level of >10 ng ml–1 of SDD and with spiked (diluted) fresh cow milk at a level >20 ng ml–1 of SDD. At these levels, the test is applicable only as a qualitative assay. The presence or absence of a test line indicates lower or higher levels of SDD, respectively. The major advantages of the one step strip test are that results can be obtained within 10 min and that all reagents are included in the test device.