Abstract
BackgroundCouples who carry alpha-thalassemia-1 deletion are at 25% risk of having a fetus with hemoglobin Bart's hydrops fetalis. Southeast Asian deletion (--(SEA)) is the most common type of alpha-thalassemia 1 among Southeast Asian populations. Thus, identification of the (--(SEA)) alpha-thalassemia 1 carrier is necessary for controlling severe alpha-thalassemia in Southeast Asian countries.ResultsUsing our generated anti zeta-globin chain monoclonal antibodies (mAbs) clones PL2 and PL3, a simple immunostick test for detecting zeta-globin chain presence in whole blood lysates was developed. The procedure of the developed immunostick test was as follows. The immunostick paddles were coated with 50 mu g/mL of mAb PL2 as capture mAb, or other control antibodies. The coated immunostick was dipped into cocktail containing tested hemolysate at dilution of 1:500, 0.25 mu g/mL biotin-labeled mAb PL3 and horseradish peroxidase-conjugated streptavidin at dilution of 1:1000. The immunostick was then dipped in precipitating substrate and the presence of zeta-globin chain in the tested sample was observed by the naked eye. Upon validation of the developed immunostick test with various types of thalassemia and normal subjects, 100% sensitivity and 82% specificity for detection of the (--(SEA)) alpha-thalassemia-1 carriers were achieved. The mAb pre-coated immunostick can be stored at room temperature for at least 20weeks.ConclusionIn this study, a novel simple immunostick test for the screening of (--(SEA)) alpha-thalassemia 1 carriers was presented. The developed immunostick test, within a single test, contains both positive and negative internal procedural controls.
