[Development of an indirect ELISA for detection of specific IgG subclasses for saffron pollen allergens]

ABSTRACT

Allergic reactions to plant pollens are a common problem. There are some evidences that saffron pollen has allergenic properties. Although IgE-mediated hypersensitivity has been reported, other immunologic mechanisms may be involved based on a variety of clinical manifestations in allergic cases. One of the mechanisms that have been suggested is specific IgG and its subclasses. In this study, an Enzyme-linked Immunosorbent Assay [ELISA] was developed for detection of specific IgG subclasses against saffron pollen extract. Briefly, microtiter plates were coated with crude extract of saffron pollen and then blocked with bovine serum albumin [BSA], and the sera were added to the plates which followed by the addition of anti-human IgG. TMB was used as substrate. Finally, after stopping the reaction with addition of HCl, plates read at 450 nm. The method was verified by testing two groups of samples, first, sera from individuals living near saffron farms in Khorasan and second, sera from individuals living in areas not having any saffron farms. The results indicated significant differences between the two groups