title: Ligase chain reaction assay for human mutations: the Sickle Cell by LCR assay.
abstract: We can detect the beta-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 oligonucleotides 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.
authors.affiliation: DNA Diagnostics Business Unit, Bio-Rad Laboratories, Hercules, CA 94547, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Reyes
authors.forename: P
authors.initials: P
authors.lastname: Carrera
authors.forename: E
authors.initials: E
authors.lastname: Cardillo
authors.forename: L
authors.initials: L
authors.lastname: Ugozzoli
authors.forename: J D
authors.initials: JD
authors.lastname: Lowery
authors.forename: C I
authors.initials: CI
authors.lastname: Lin
authors.forename: M
authors.initials: M
authors.lastname: Go
authors.forename: M
authors.initials: M
authors.lastname: Ferrari
authors.forename: R B
authors.initials: RB
authors.lastname: Wallace
mesh_terms: D000483:Alleles; D000755:Anemia, Sickle Cell / Q000235:genetics*; D003124:Colorimetry; D004247:DNA / Q000032:analysis* / Q000235:genetics; D011088:DNA Ligases / Q000378:metabolism*; D005838:Genotype; D005914:Globins / Q000235:genetics*; D006451:Hemoglobin, Sickle / Q000235:genetics; D019382:Human Growth Hormone / Q000235:genetics; D006801:Humans; D009154:Mutation; D009841:Oligonucleotides
chemical_list: D006451:Hemoglobin, Sickle; D009841:Oligonucleotides; D019382:Human Growth Hormone; D005914:Globins; D004247:DNA; D011088:DNA Ligases