Full validation of an ELISA for quantification of antibodies against Haemophilus influenzaetype b capsular polysaccharide

Since 1981, ELISA methods have been used for the quantification of antibodies against Haemophilus influenzae type b (Hib), but their complete validation has been never reported. This work described the full validation of an ELISA for the quantification of antibodies against Hib capsular polysaccharide, following eleven parameters recommended by European Medicines Agency and FDA guidelines. The method satisfied the acceptance criteria for the required parameters. The use of percent of inhibition and parallelism fulfillment as acceptance criteria for specificity was discussed. This ELISA will be a reliable device for the evaluation of Hib-vaccination efficacious.d