Molecular genetic diagnosis of sickle cell disease using dried blood specimens on blotters used for newborn screening

SummaryThe 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.

[1]  C. Oste,et al.  Polymerase Chain Reaction , 2002 .

[2]  A. Dilella,et al.  Correlation between polymorphic DNA haplotypes at phenylalanine hydroxylase locus and clinical phenotypes of phenylketonuria. , 1987, The Journal of pediatrics.

[3]  A. Covone,et al.  TROPHOBLAST CELLS IN PERIPHERAL BLOOD FROM PREGNANT WOMEN , 1984, The Lancet.

[4]  Henry A. Erlich,et al.  Enzymatic amplification of ?-globin genomic sequences and restriction site analysis for diagnosis of , 1985 .

[5]  U Landegren,et al.  A ligase-mediated gene detection technique. , 1988, Science.

[6]  J. Gitschier,et al.  An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. , 1987, The New England journal of medicine.

[7]  K. Itakura,et al.  Detection of sickle cell beta S-globin allele by hybridization with synthetic oligonucleotides. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. McCabe,et al.  DNA microextraction from dried blood spots on filter paper blotters: potential applications to newborn screening , 1987, Human Genetics.

[9]  R. Teplitz,et al.  Discrimination among the human beta A, beta S, and beta C-globin genes using allele-specific oligonucleotide hybridization probes. , 1985, American journal of human genetics.

[10]  X. Estivill,et al.  A candidate for the cystic fibrosis locus isolated by selection for methylation-free islands , 1987, Nature.

[11]  A. Dilella,et al.  SCREENING FOR PHENYLKETONURIA MUTATIONS BY DNA AMPLIFICATION WITH THE POLYMERASE CHAIN REACTION , 1988, The Lancet.

[12]  J Gitschier,et al.  An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. , 1987, The New England journal of medicine.

[13]  Henry A. Erlich,et al.  Characterization of β-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA , 1987, Nature.

[14]  A. Christiano,et al.  Rapid prenatal diagnosis of sickle cell diseases using oligonucleotide ligation assay coupled with laser‐induced capillary fluorescence detection , 2002, Prenatal diagnosis.

[15]  W. Rosse,et al.  The cooperative study of sickle cell disease: review of study design and objectives. , 1982, The American journal of pediatric hematology/oncology.

[16]  R. Nussbaum,et al.  Newborn screening for sickling hemoglobinopathies. Houston, 1976 to 1980. , 1984, American journal of diseases of children.

[17]  P Berg,et al.  Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. , 1977, Journal of molecular biology.

[18]  K. Mullis,et al.  Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. , 1988, Science.

[19]  C. Levenson,et al.  Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes. , 1988, The New England journal of medicine.

[20]  R. Schabel.  Diagnosis of sickle-cell anemia in the prosthetic patient. , 1968, The Journal of prosthetic dentistry.

[21]  R. Williamson,et al.  GUTHRIE SPOTS FOR DNA-BASED CARRIER TESTING IN CYSTIC FIBROSIS , 1988, The Lancet.

[22]  Henry A. Erlich,et al.  The polymerase chain reaction. , 1989, Trends in genetics : TIG.

[23]  E Vichinsky,et al.  Prophylaxis with oral penicillin in children with sickle cell anemia. A randomized trial. , 1986, The New England journal of medicine.

[24]  David J. Werrett,et al.  Forensic application of DNA ‘fingerprints’ , 1985, Nature.

[25]  C. Caskey.  Disease diagnosis by recombinant DNA methods. , 1987, Science.

[26]  H A Erlich,et al.  Rapid prenatal diagnosis of sickle cell anemia by a new method of DNA analysis. , 1987, The New England journal of medicine.

[27]  C. Frömmel.  Newborn Screening for Sickle Cell Disease and Other Hemoglobinopathies: A Short Review on Classical Laboratory Methods—Isoelectric Focusing, HPLC, and Capillary Electrophoresis , 2018, International journal of neonatal screening.

[28]  K. Mullis,et al.  Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. , 1985, Science.

[29]  A. Munnich,et al.  GUTHRIE CARDS FOR DETECTION OF POINT MUTATIONS IN PHENYLKETONURIA , 1988, The Lancet.