title: Accurate and rapid detection of heterozygous carriers of a deletion by combined polymerase chain reaction and high-performance liquid chromatography.
abstract: We have developed a technique to detect accurately heterozygous carriers of a deletion. Specific target sequences were amplified by the polymerase chain reaction (PCR), and the products subsequently were analyzed by high-performance liquid chromatography. Examples from four loci demonstrated that 24-27 cycles of amplification for a single-copy DNA, based on 50 ng of genomic DNA, results in excellent quantitation that readily permits the detection of heterozygous carriers of a deletion. We have demonstrated that triplex PCR (three targets in a single PCR) entails no loss of precision. We also have demonstrated that this method can accurately differentiate the heterozygous carriers of a deletion from normal individuals in four family studies, three for Duchenne muscular dystrophy patients and one for a hemophilia B patient.
authors.affiliation: Department of Genetics, Radiation Effects Research Foundation, Hiroshima, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Asakawa
authors.forename: C
authors.initials: C
authors.lastname: Satoh
authors.forename: Y
authors.initials: Y
authors.lastname: Yamasaki
authors.forename: S H
authors.initials: SH
authors.lastname: Chen
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D006580:Genetic Carrier Screening; D006242:Haptoglobins / Q000235:genetics*; D006454:Hemoglobins / Q000235:genetics*; D006467:Hemophilia A / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D006242:Haptoglobins; D006454:Hemoglobins; D009838:Oligodeoxyribonucleotides; D004247:DNA