Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes

N Engl J Med. 1988 Sep 1;319(9):537-41. doi: 10.1056/NEJM198809013190903.

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.

MeSH terms

  • Alleles
  • Anemia, Sickle Cell / diagnosis*
  • Colorimetry
  • DNA / analysis*
  • DNA-Directed DNA Polymerase / pharmacology
  • Female
  • Fetal Diseases / diagnosis*
  • Gene Amplification*
  • Globins / genetics*
  • Horseradish Peroxidase
  • Humans
  • Nucleic Acid Hybridization
  • Oligonucleotides*
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • Retrospective Studies
  • Thalassemia / diagnosis*

Substances

  • Oligonucleotides
  • Globins
  • DNA
  • Horseradish Peroxidase
  • DNA-Directed DNA Polymerase