Summary
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 μl of whole blood. We utilized the DNA from a 1/4 inch diameter specimen (12 μl equivalent) for polymerase chain reaction amplification of the β-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.
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References
Caskey CT (1987) Disease diagnosis by recombinant DNA methods. Science 236:1223–1229
Conner BJ, Reyes AA, Morin C, Itakura K, Teplitz RL, Wallace RB (1983) Detection of sickle cell βS-globin allele by hybridization with synthetic oligonucleotides. Proc Natl Acad Sci USA 80:278–282
Covone AE, Mutton D, Johnson PM, Adinolfi M (1984) Trophoblast cells in the peripheral blood from pregnant women. Lancet II: 841–843
DiLella AG, Huang W-M, Woo SLC (1988) Screening for phenylketonuria mutations by DNA amplification with the polymerase chain reaction. Lancet I:497–499
Embury SH, Scharf SJ, Saiki RK, Gholson MA, Golbus M, Arnheim N, Erlich HA (1987) Rapid prenatal diagnosis of sickle cell anemia by a new method of DNA analysis. N Engl J Med 316: 656–661
Estivill X, Farrall M, Scambler PJ, Bell GM, Hawley KMF, Lench NJ, Bates GP, Kruyer HC, Frederick PA, Stanier P, Watson EK, Williamson R, Wainwright BJ (1987) A candidate for the cystic fibrosis locus isolated by selection for methylation-free islands. Nature 326:840–845
Garrick MD (1987) Technical options for screening newborns for hemoglobinopathies. In: Therrel BL (ed) Advances in neonatal screening. Elsevier, Amsterdam, pp 417–423
Gaston M, Rosse WF (1982) The cooperative study of sickle cell disease: review of study design and objectives. Am J Pediatr Hematol Oncol 4:197–201
Gaston MH, Verter JI, Woods G, Pegelow C, Kelleher J, Presbury G, Zarkowsky H, Vichinsky E, Iyer R, Lobel JS, Diamond S, Holbrook CT, Gill FM, Ritchey K, Falletta JM, Prophylactic Penicillin Study Group (1986) Prophylaxis with oral penicillin in children with sickle cell anemia — a randomized trial. N Engl J Med 314:1593–1599
Gill P, Jeffreys AJ, Werrett DJ (1985) Forensic application of DNA “fingerprints”. Nature 318:577–579
Guttler F, Lidsky AS, Ledley FD, DiLella AG, Sullivan SC, Woo SLC (1987) Correlation between polymorphic DNA haplotypes at the phenylalanine hydroxylase locus and the clinical phenotypes of phenylketonuria. J Pediatr 110:68–71
Kogan SC, Doherty M, Gitschier J (1987) An improved method for prenatal diagnosis of genetic diseases by analysis of amplified sequences: application to hemophilia A. N Engl J Med 317:985–990
Landegren U, Kaiser R, Sanders J, Hood L (1988) A ligase-mediated gene detection technique. Science 241:1077–1080
Lyonnet S, Caillaud C, Rey F, Berthelon M, Frezal J, Rey J, Munnich A (1988) Guthrie cards for detection of point mutations in phenylketonuria. Lancet II:507
McCabe ERB, Huang S-Z, Seltzer WK, Law ML (1987) DNA micro-extraction from dried blood spots on filter paper blotters: potential applications to newborn screening. Hum Genet 75:213–216
National Committee for Clinical Laboratory Standards (1985) Blood collection on filter paper for neonatal screening programs; tentative standards. (NCCLS publication LA4-T) NCCLS, Villanova, Pa
Nussbaum RL, Powell C, Graham HL, Caskey CT, Fernbach DJ (1984) Newborn screening for sickling hemoglobinopathies —Houston 1976 to 1980. Am J Dis Child 138:44–48
Oste C (1988) Polymerase chain reaction. BioTech 6:162–167
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230:1350–1354
Saiki RK, Chang CA, Levenson CH, Warren TC, Boehm CD, Kazazian HH Jr, Erlich HA (1988a) Diagnosis of sickle cell anemia and β-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes. N Engl J Med 319: 537–541
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988b) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Schneider RG (1986) Laboratory identification of hemoglobin variants in the newborn. In: Carter TP, Willey AM (eds) Genetic disease — screening and management. Liss, New York, pp 137–150
Studencki AB, Conner BJ, Impraim LC, Teplitz RL, Wallace RB (1985) Discrimination among the human βA, βS, βC-globin genes using allele-specific oligonucleotide hybridization probes. Am J Hum Genet 37:42–51
Wethers DL, Panel (1987) Newborn screening for sickle cell disease and other hemoglobinopathies. National Institutes of Health Consensus Development Conference Statement. 6(9):1–22
Williams C, Weber L, Williamson R, Hjelm M (1988) Guthrie spots for DNA-based carrier testing in cystic fibrosis. Lancet II:693
Wong C, Dowling CE, Saiki RK, Higuchi RG, Erlich HA, Kazazian HH Jr (1987) Characterization of β-thalassemia mutations using direct genomic sequencing of amplified single copy DNA. Nature 330:384–386
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Jinks, D.C., Minter, M., Tarver, D.A. et al. Molecular genetic diagnosis of sickle cell disease using dried blood specimens on blotters used for newborn screening. Hum Genet 81, 363–366 (1989). https://doi.org/10.1007/BF00283692
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DOI: https://doi.org/10.1007/BF00283692