RT Journal Article
SR Electronic
T1 A Method of HbF Determination for Potential Use in Underdeveloped Countries
JF American Society for Clinical Laboratory Science
JO Clin Lab Sci
FD American Society of Chemistry and Laboratory Science
SP 212
OP 218
DO 10.29074/ascls.25.4.212
VO 25
IS 4
A1 Schumacher, Sarah N.
A1 Randolph, Tim R.
YR 2012
UL http://hwmaint.clsjournal.ascls.org/content/25/4/212.abstract
AB The objective of this study was to develop a simple, cost-effective method of HbF determination potentially useable in underdeveloped countries to determine sickle cell patient response to hydroxyurea treatment. Normal adult blood (HbA), cord blood (HbF), and a 50:50 mixture (HbA+F) were the three sample types used in procedure development. Normal blood samples were collected from the research team, and de-identified cord blood samples were provided by Cardinal Glennon Pediatric Research Institute, St. Louis, MO. The hematocrit of all blood samples was standardized to 35%. The method, based on the Kleihauer-Betke (K-B) test principle, used a citrate solution to selectively elute HbA from RBCs while HbF remained intracellular, and spectrophotometric absorbance of the eluate was the primary outcome measure. A procedure was developed and optimized utilizing a 395 nm wavelength, 30 sec centrifugation time, 6 min incubation time, 20 μL blood volume, and 0.07 M sodium citrate in a 0.06 M sodium phosphate buffer solution. Reproducibility was demonstrated (N = 39) with a mean HbA absorbance of 1.285 (SD 0.069), mean HbA+F absorbance of 0.690 (SD 0.050), and mean HbF absorbance of 0.035 (SD 0.005), also exhibiting linearity (r2 = 0.99). This simple, cost-effective method of HbF determination shows potential as a basis for determining sickle cell patient response to hydroxyurea treatment in underdeveloped countries.ABBREVIATIONSHbS-sickle hemoglobin, HbA-normal hemoglobin, SCD-sickle cell disease, SCT-sickle cell trait, RBCs-red blood cells, HPLC-high performance liquid chromategraphy, HbF-fetal hemoglobin, K-B-Kleihauer-Betke.