| Mycotoxins are secondary metabolites produced by certain fungi that can grow on foods such as cereals, dried fruits, spices, legumes and others under certain environmental conditions. Ochratoxin A(OTA), is one of the most important mycotoxins with worldwide occurrence and toxicity, together with aflatoxins and zearalenone, which brought bad effect on human and animal’s health. Existing testing methods such as HPLC and GC-MS were accurate, However, And all of them are too expensive instruments to satiation the requirement of witnessed inspections for OTA. So to built a rapid and convenient detection method for OTA analysis is absolutely desirable.In this study, the OTA was conjugated to a carrier protein (BSA or OVA) by the active-ester method. The spleen cells from BALB/c immunized with OTA-BSA were fused with SP2/0myeloma cells and a fused cell line, which secreted stable McAb against OTA was screened with a indirect ELISA. The hybridoma were cloned and subcloned by limited dilution method. The titer of McAb in astices was1:204800. McAb has specificity integrated with OTA conjugated antigen that determined by gold-immunochromatography assay (GICA) which showed good sensitivity with an IC50of0.5ng/mL for detection of OTA and had not cross reactivity with OTB, OTC, and coumadin.Colloidal gold, of which the diameter is about25nm, was obtained by reducing the gold chloride with sodium citrate and labeled with anti-OTA McAb as detection reagent. The optimum pH for labelling was about8.5and the amount of mAb was18μg/mL. The colloidal gold-labeled mAb was stabilized by1%BSA. The detection sensitivity of OTA-kit was5ng/mL. |
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