Reverse dot-blot detection of the African-American β-thalassemia mutations

P. Sutcharitchan, R. Saiki, T. H.J. Huisman, A. Kutlar, V. McKie, H. Erlich, S. H. Embury

Research output: Contribution to journalArticlepeer-review

42 Scopus citations


DNA-based diagnosis of the β thalassemias provides accuracy to newborn screening, genetic counseling, and prenatal diagnosis. However, the use of polymerase chain reaction (PCR)-based methods is challenged by the great number of different β-thalassemia mutations that exist even within defined ethnic groups. In this regard, the reverse dot-blot method offers a means of screening for several mutations with a single hybridization reaction. We have applied the reverse dot-blot method to the detection of the β-thalassemia mutations of African-Americans. We used two biotin-labeled primer pairs in a duplex reaction to amplify and label two β-globin target DNA fragments that encompass all known African-American β-thalassemia mutations. The PCR products were denatured and hybridized to polyT-tailed, membrane-fixed, allele-specific probe pairs for the hemoglobin (Hb) S, Hb C, and 14 β- thalassemia mutations and their corresponding wild-type sequences. Seven common mutations plus Hb S and Hb C were included on one diagnostic strip, and seven less common β-thalassemia mutations were included on another strip. Carefully controlled, high stringency hybridization allowed accurate distinction of these alleles. Reverse dot-blot diagnosis of the less common β-thalassemia mutations precludes the need for alternative, more technically challenging methods. This method provides a rapid, accurate method for diagnosis of β thalassemia among African-Americans and other ethnic groups in which β thalassemia occurs.

Original languageEnglish (US)
Pages (from-to)1580-1585
Number of pages6
Issue number4
StatePublished - Aug 15 1995

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology


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