The last 25 years dried blood spots are used for the diagnosis of hereditary diseases as hyperthyroidism, phenylketonuria and enzymopathies. Today dried blood spots are used as well for the screening of hemoglobinopathies and the structural characterization of abnormal hemoglobin variants at protein and gene level. In the case of hemoglobin, a small piece of the blood spot is cut and analyses performed by electrofocusing, either on agarose, or on polyacrylamide gels and by anion or cation exchange HPLC. These methods are suitable for the diagnosis of abnormal hemoglobin variants, mainly of HbS, but their efficacy in the case of thalassemias is limited.

The aim of this communication is to evaluate a cation excange HPLC based method for the screening of hemoglobinopathies, thalassemias and diabetes from blood spots collected on humidified filter paper. Fifty random blood specimens were used in this study. Blood was collected in K3EDTA vials, while blood drops (60-80μl) were applied on filter papers. Filter papers were dipped in H2O, placed in plastic bags and stored at 4 °C until used. Elution from the paper was performed by the addition of 400μl solution containing 7mM KCN, 24mM potassium phthalate and 2ml/l saponin pH 5.5. The hemolysates were prepared in 1/20 dilution in H2O. Cation exchange HPLC analysis was performed by the Variant Bio-Rad system, using the “thalassemia short” program.

The main results showed a significant correlation between hemoglobin values observed in liquid and paper sampling (HbA2 r=0.977, HbF r=0.999, HbA1c r=0.954, HbA1d r=0.884).

The described methodology has the following advantages: is not aggressive for the population tested; the sample collection, handling, storage and shipment are considerably easy; it is suitable for screening test of large populations as students, soldiers etc. and characterized by accuracy and reproducibility. In our opinion, these preliminary results lead to the conclusion that this methodology can constitute a national pilot program for screening thalassemias, hemoglobinopathies and diabetes.