Simultaneous measurement of HbA1C and Hemoglobin by Turbidimetric inhibition immunoassay from a single punch of dried blood spot samples

Abstract

Hemoglobin and glycosylated hemoglobin (HbA1C) are frequently monitored health indicators in population based studies for information about the status of nutrition and diabetes control. We present here possibly for the first time the findings of simultaneous estimation of Hemoglobin and HbA1C on Dried blood spot (DBS) samples by a single test. Validation was done by turbidimetric inhibition immunoassay (TINIA) using Roche Integra 400 plus instrument. Paired whole blood and DBS samples were tested for HbA1C estimation by Integra 400 plus. Total hemoglobin values obtained during HbA1C estimation were compared with hemoglobin values estimated by Coulter AcT 5 Diff CP Hematology counter. Agreement in HbA1C and hemoglobin values between paired whole blood and DBS samples was found to be high with R2 values of 0.994 and 0.9349, respectively. Intra- and inter- assay precision was found to be within 10% for both parameters. Values obtained after assaying DBS samples prepared by spotting proficiency samples on Whatman 903 protein saver cards demonstrated acceptable standard deviation indices resulting in successful participation in EQAS programs for both these parameters. The results reveal the potential of TINIA for simultaneous estimation of hemoglobin and HbA1C from a single punch of the DBS samples.