title: Prenatal diagnosis by enzymatic amplification and restriction endonuclease digestion for detection of haemoglobins A, S and C.
abstract: The detection of the single base pair mutations at codon 6 of the beta-globin gene is important for the prenatal diagnosis of sickle-cell anaemia and SC disease. A novel procedure has been designed to create a restriction site at both the beta A and beta C alleles to facilitate the discrimination of haemoglobins A, S and C. The general principle of this procedure is to enzymatically amplify genomic DNA using a modified primer containing an altered 3'-terminal nucleotide to create these restriction sites. After this modified primer has been efficiently incorporated into amplified DNA, the PCR products are digested with the restriction enzymes Ava I and Sty I. Ava I recognizes a site in amplified DNA containing a beta A allele, and Sty I recognizes a site in DNA containing a beta C allele. Since the beta A and beta C alleles can be distinguished directly by the presence of a restriction site, the beta S allele can be identified indirectly. All three beta-globin alleles are easily distinguished by size and pattern of electrophoresed fragments on agarose gels. This procedure is specific and sensitive, thus permitting rapid, economical diagnosis of sickle-cell anaemia and SC disease.
authors.affiliation: Department of Pathology, University of California, Davis Medical Center 95816.
authors.forename: S L
authors.initials: SL
authors.lastname: Hatcher
authors.forename: Q T
authors.initials: QT
authors.lastname: Trang
authors.forename: K M
authors.initials: KM
authors.lastname: Robb
authors.forename: R L
authors.initials: RL
authors.lastname: Teplitz
authors.forename: J R
authors.initials: JR
authors.lastname: Carlson
mesh_terms: D000755:Anemia, Sickle Cell / Q000175:diagnosis*; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D005260:Female; D006441:Hemoglobin A / Q000235:genetics*; D006444:Hemoglobin C / Q000235:genetics*; D006450:Hemoglobin SC Disease / Q000175:diagnosis; D006451:Hemoglobin, Sickle / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*
chemical_list: D006451:Hemoglobin, Sickle; D004247:DNA; D006444:Hemoglobin C; D006441:Hemoglobin A; D004262:DNA Restriction Enzymes