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[Quantification of total IgE on microtiter plates by enzyme immunoassay]
Abstract:
This work describes an immunoenzymatic method for the detection of total IgE in human sera. The assay is performed using specially formulated microtiter plates as solid phase (Dynatech microElisa). This facilitates the dispensing of the reagents, the washing steps and the reading of the results. The whole assay is done using commercially available reagents. This guarantees the high specificity and reliability of the test as well as the low interassay variation. The method uses 10 micro liters of human serum and is performed in two work days. IgE levels ranging between 10 and 1000 I.U./ml can be accurately assayed without modifications of the test volumes. The microplate test, when compared with other methods, has an additional advantage: the absorbance values of the wells can be processed by a microcomputer. The method has been compared with a commercial kit (EnzygnostTM) and the correlation found in 150 cases was excellent (r = 0.983). Using this method, we have assayed more than one thousand sera with very satisfactory results. In addition, the cost of each test has been reduced by 75%.