Multivariant confirmation of sickle cell disease using a non- radioactive minisequencing reaction

T. Tuuminen, H. Ingman, B. L. Therrell, A. Kallio

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


A non-radioactive solid-phase minisequencing method for confirmation of abnormal hemoglobin variants causing sickle cell disease has been developed. In this method amplified 5'-biotinylated target sequences containing normal and mutation sites are immobilized onto streptavidin-coated microplates. Detection primers corresponding to target sequences are annealed immediately adjacent to the mutation site and single-step, haptenlabeled nucleotide primer extension reactions are performed. The incorporation of the labeled nucleotide is detected through immunological reaction with an enzyme-labeled anti-hapten conjugate and a substrate. The method enables confirmation of mutations of the β-globin gene variants (Hbs S, C, E, D-Punjab, O-Arab) and the α-globin gene variant (Hb G-Philadelphia). The test was evaluated using characterized dried blood spot specimens (n = 100). The advantages of the procedure are easy performance and objectiveness. The non-radioactive minisequencing assay will prove helpful for genotyping in neonatal screening for hemoglobinopathies and in prenatal and pre-implantational diagnostics.

Original languageEnglish (US)
Pages (from-to)71-89
Number of pages19
Issue number1
StatePublished - 1997
Externally publishedYes

ASJC Scopus subject areas

  • Hematology
  • Clinical Biochemistry
  • Genetics(clinical)
  • Biochemistry, medical


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