title: Enzyme enhanced quantitative determination of multiple DNA targets based on capillary electrophoresis.
abstract: In the current paper, enzyme enhanced simultaneous quantitative determination of multiple DNA targets based on capillary electrophoresis (CE) was described. We used three biotin-modified DNA probes, which reacted with avidin-conjugated horseradish peroxidase (avidin-HRP) conjugate to obtain the HRP labeled probes, to hybridize with three corresponding targets. The resulting mixture containing double-strand DNA (dsDNA)-HRP, excess single-strand DNA (ssDNA)-HRP and remaining avidin-HRP was separated by capillary electrophoresis, and then the system of HRP catalyzing H(2)O(2)/o-aminophenol (OAP) reaction was adopted. The catalytic product was detected with electrochemical detection. With this protocol, the limits of quantification for the hybridization assay of 21-, 39- and 80-mer DNA fragments were of 1.2 x 10(-11), 2.4 x 10(-11) and 3.0 x 10(-11)M, respectively. The multiplex assay also provided good specificity without any cross-reaction.
authors.affiliation: Department of Chemistry, Nanjing University, Nanjing, China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Li
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Zhan
authors.forename: Shusheng
authors.initials: S
authors.lastname: Zhang
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Chen
mesh_terms: D001360:Avidin / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism; D004247:DNA / Q000032:analysis* / Q000378:metabolism*; D055664:Electrochemical Techniques / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C083986:avidin-horseradish peroxidase complex; D001360:Avidin; D001710:Biotin; D004247:DNA; D006735:Horseradish Peroxidase