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Clinical Chemistry 42: 704-710, 1996;
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Clinical Chemistry, Vol 42, 704-710, Copyright © 1996 by American Association for Clinical Chemistry

Automated HPLC screening of newborns for sickle cell anemia and other hemoglobinopathies

JW Eastman, R Wong, CL Liao and DR Morales
California Department of Health Services, Genetic Disease Laboratory, Berkeley, CA 94710, USA.

Automated HPLC is used to test dried blood-spot specimens from newborns for hemoglobins (Hb) F, A, S, C, E, and D. We present the method and report on its performance determined during >4 years of testing 2.5 x 10(6) newborns. The method features automated derivation of presumptive phenotypes; quantitative quality control and proficiency testing; throughput of one specimen per minute; small sample volume; hemoglobin concentrations quantified with an interlaboratory CV of 14-18%; retention times with interlaboratory CV of <2% and matching, within +/- 0.03 min, of laboratories and reagent lots; control of peak resolution; 0.5% detection limit for Hb S and C, and 1.0% for Hb F, A, E, and D; few interferences; and negligible background and carryover. Shortcomings of the method are the absence of microplate barcode identification and the need for manually pipetting the sample eluate into the microplate.


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Copyright © 1996 by the American Association for Clinical Chemistry.