title: An assay of gene copy number and its application based on heteroduplex products.
abstract: The aim of this study was to set up a simple and efficient method for detecting gene copy number, based on heteroduplex products from single-tube PCR/DHPLC. Single-nucleotide polymorphisms (SNPs) on the α-globin gene and chromosome 21 were used as examples. And the formula for quantitative calculation of gene copy number was deduced-based on the peak heights of homoduplexes and heteroduplexes on the DHPLC pattern. 27 samples (14 normal DNA and 13 cases of trisomy-21) were assessed with this method, and 160 samples (48 normal DNA and 112 α-thalassemia samples) were assessed with this method combined with a duplex PCR/DHPLC. Results for 184 of 187 cases were concordant with the known genotypes; three cases of trisomy-21 could not be detected because the target SNPs were homozygous. In conclusion, quantitative assessment of heteroduplex products from single-tube PCR/DHPLC is simple and rapid, and can be used to detect α-thalassemia gene deletions (α(-3.7), α(-4.2)) and trisomy-21.
authors.affiliation: Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. jiaxy@hotmail.com
authors.forename: Xingyuan
authors.initials: X
authors.lastname: Jia
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Wei
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Meiting
authors.initials: M
authors.lastname: Cao
authors.forename: Lirong
authors.initials: L
authors.lastname: Wang
authors.forename: Meiling
authors.initials: M
authors.lastname: Zheng
authors.forename: Jingzhong
authors.initials: J
authors.lastname: Liu
mesh_terms: D000510:Alpha-Globulins / Q000235:genetics; D001681:Biological Assay / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002891:Chromosomes, Human, Pair 21; D004252:DNA Mutational Analysis / Q000379:methods; D004314:Down Syndrome / Q000175:diagnosis / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage / Q000235:genetics*; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017085:alpha-Thalassemia / Q000175:diagnosis / Q000235:genetics
chemical_list: D000510:Alpha-Globulins; D009692:Nucleic Acid Heteroduplexes