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

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

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

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
JournalBlood
Volume86
Issue number4
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Thalassemia
African Americans
Hemoglobin C
Sickle Hemoglobin
Mutation
Polymerase chain reaction
Screening
Globins
DNA
Biotin
Reaction products
Ethnic Groups
Labels
Membranes
Alleles
Polymerase Chain Reaction
Genetic Counseling
Prenatal Diagnosis
Newborn Infant

ASJC Scopus subject areas

  • Immunology
  • Biochemistry
  • Hematology
  • Cell Biology

Cite this

Sutcharitchan, P., Saiki, R., Huisman, T. H. J., Kutlar, A., McKie, V., Erlich, H., & Embury, S. H. (1995). Reverse dot-blot detection of the African-American β-thalassemia mutations. Blood, 86(4), 1580-1585.

Reverse dot-blot detection of the African-American β-thalassemia mutations. / Sutcharitchan, P.; Saiki, R.; Huisman, T. H.J.; Kutlar, Abdullah; McKie, V.; Erlich, H.; Embury, S. H.

In: Blood, Vol. 86, No. 4, 01.01.1995, p. 1580-1585.

Research output: Contribution to journalArticle

Sutcharitchan, P, Saiki, R, Huisman, THJ, Kutlar, A, McKie, V, Erlich, H & Embury, SH 1995, 'Reverse dot-blot detection of the African-American β-thalassemia mutations', Blood, vol. 86, no. 4, pp. 1580-1585.
Sutcharitchan P, Saiki R, Huisman THJ, Kutlar A, McKie V, Erlich H et al. Reverse dot-blot detection of the African-American β-thalassemia mutations. Blood. 1995 Jan 1;86(4):1580-1585.
Sutcharitchan, P. ; Saiki, R. ; Huisman, T. H.J. ; Kutlar, Abdullah ; McKie, V. ; Erlich, H. ; Embury, S. H. / Reverse dot-blot detection of the African-American β-thalassemia mutations. In: Blood. 1995 ; Vol. 86, No. 4. pp. 1580-1585.
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