Ligase chain reaction assay for human mutations: The sickle cell by LCR assay

Antonio A. Reyes, Paola Carrera, Elena Cardillo, Luis Ugozzoli, Jlmmie D. Lowery, Ching I P Lin, Matthew Go, Maurizio Ferrari, R. Bruce Wallace

Research output: Contribution to journalArticle

Abstract

We can detect the β-globin gene sickle cell mutation by using an assay based on the ligase chain reaction. The simultaneous amplification of the human growth hormone gene in the same reaction serves as a control for the amount of template DNA or amplification efficiency. Ligation products, which are biotinylated at one end and tagged with an arbitrary 'tail' sequence at the other, are captured by hybridization to 'tail'-complementary oligonudeotides immobilized on polystyrene microwells. The captured ligation products are detected colorimetrically by use of streptavidin-alkaline phosphatase conjugate. In a study of 24 subjects, the assay unequivocally discriminated among normal, carrier, and sickle cell genotypes.

Original languageEnglish
Pages (from-to)40-44
Number of pages5
JournalClinical Chemistry
Volume43
Issue number1
Publication statusPublished - 1997

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Keywords

  • biotin-streptavidin interaction
  • DNA amplification
  • genetic diseases
  • hemoglobinopathies
  • human growth hormone gene

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Reyes, A. A., Carrera, P., Cardillo, E., Ugozzoli, L., Lowery, J. D., Lin, C. I. P., Go, M., Ferrari, M., & Wallace, R. B. (1997). Ligase chain reaction assay for human mutations: The sickle cell by LCR assay. Clinical Chemistry, 43(1), 40-44.