Automated Dithionite Test for Rapid, Inexpensive Detection of Hemoglobin S and Non-S Sickling Hemoglobinopathies

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Abstract
Automated adaptations of dithionite and urea—dithionite tube tests are accurate, reliable, inexpensive methods for detecting hemoglobin S. More than 3,000 individuals have been screened (120 determinations per hour; reagent cost, 2 to 4 cents per test). The dithionite reagent consists of potassium phosphate, sodium dithionite, and saponin. When S hemoglobin contacts this reagent, the red cells lyse, and the hemoglobin deoxygenates and sickles, forming a hydrophobic-bond-dependent nematic liquid crystal system that is manifested as turbidity. The resulting AutoAnalyzer curve is strikingly and diagnostically different from that produced by hemoglobin A in the same reagent. Specificity of the automated dithionite test may be enhanced by use of the automated urea—dithionite test, which consists of a specimen set of two aliquots: one traverses a dithionite line, the other a urea—dithionite line. A comparison of transmittance in the two lines yields typical diagnostic curves because the urea disperses the sickling, with a consequently increased transmittance over that of the dithionite aliquot. Methods are discussed for recognizing non-S sickling hemoglobins and a few other rare hemoglobinopathies.

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