title: Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes.
abstract: We have developed a simple and rapid nonradioactive method for detecting genetic variation and have applied it to the diagnosis of sickle cell anemia and beta-thalassemia. The procedure involves the selective amplification of a segment of the human beta-globin gene with oligonucleotide primers and a thermostable DNA polymerase, followed by hybridization of the amplified DNA with allele-specific oligonucleotide probes covalently labeled with horseradish peroxidase. The hybridized probes were detected with a simple colorimetric assay. We demonstrated the usefulness of this method in a retrospective analysis of two pregnancies at risk for beta-thalassemia and one at risk for sickle cell anemia, as well as in an analysis of nine DNA samples simulating three family sets.
authors.affiliation: Department of Human Genetics, Cetus Corporation, Emeryville, Calif. 94608.
authors.forename: R K
authors.initials: RK
authors.lastname: Saiki
authors.forename: C A
authors.initials: CA
authors.lastname: Chang
authors.forename: C H
authors.initials: CH
authors.lastname: Levenson
authors.forename: T C
authors.initials: TC
authors.lastname: Warren
authors.forename: C D
authors.initials: CD
authors.lastname: Boehm
authors.forename: H H
authors.initials: HH
authors.lastname: Kazazian
authors.forename: H A
authors.initials: HA
authors.lastname: Erlich
mesh_terms: D000483:Alleles; D000755:Anemia, Sickle Cell / Q000175:diagnosis*; D003124:Colorimetry; D004247:DNA / Q000032:analysis*; D004259:DNA-Directed DNA Polymerase / Q000494:pharmacology; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis*; D005784:Gene Amplification; D005914:Globins / Q000235:genetics*; D006735:Horseradish Peroxidase; D006801:Humans; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012189:Retrospective Studies; D013789:Thalassemia / Q000175:diagnosis*
chemical_list: D009841:Oligonucleotides; D005914:Globins; D004247:DNA; D006735:Horseradish Peroxidase; D004259:DNA-Directed DNA Polymerase