Molecular genetic diagnosis of sickle cell disease using dried blood specimens on blotters used for newborn screening.
The protein-based technologies used to screen newborns for sickle cell disease require confirmation with a liquid blood specimen. We have developed a strategy for rapid and specific genotypic diagnosis using DNA extracted from a dried blood spot on the filter paper blotter used to screen newborns. DNA could be microextracted from a specimen as small as a 1/8 inch diameter punched disc representing the dried equivalent of approximately 3 microliters of whole blood. We utilized the DNA from a 1/4 inch diameter specimen (12 microliters equivalent) for polymerase chain reaction amplification of the beta-globin region spanning the sickle cell mutation with detection by allele-specific oligonucleotide probes. Molecular confirmation of genotype from the original blotter would reduce the personnel costs associated with obtaining follow-up liquid blood specimens and would provide information to the family in a more timely and less equivocal manner.
Jinks
D C
DC
Department of Microbiology, State University of New York, Buffalo 14214.
Minter
M
M
Tarver
D A
DA
Vanderford
M
M
Hejtmancik
J F
JF
McCabe
E R
ER
D000755:Anemia, Sickle Cell / Q000175:diagnosis / Q000235:genetics*; D001800:Blood Specimen Collection; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis*; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D007231:Infant, Newborn
D004247:DNA