title: Chemiluminescence analysis for HBV-DNA hybridization detection with magnetic nanoparticles based DNA extraction from positive whole blood samples.
abstract: Molecular detection of HBV has a significant impact on prognosis and therapy of the disease. In this paper, a sensitive nucleic acid detection method of HBV was established taking advantage of magnetic nanoparticles (MNPs), chemiluminescence (CL) and polymerase chain reaction (PCR). HBV-DNA was extracted from hepatitis B positive human blood samples using MNPs adsorption method and biotin was labeled on the DNA segment after base insertion of bintin-dUTP in PCR. The biotinylated DNA segment was captured by amino probe immobilized on carboxyl MNPs and was detected by the chemiluminescence system of alkaline phosphatase catalyzing 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy) phenyl-1, 2-dioxetane. Different concentrations of HBV-DNA were detected under the optimized experiment conditions and the relevant CL intensity were obtained, which provided a novel research or clinic diagnosis method for the quantification detection of HBV-DNA.
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, P. R. China.
authors.forename: Nongyue
authors.initials: N
authors.lastname: He
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
authors.forename: Chuanyan
authors.initials: C
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Zeng
authors.forename: Yan
authors.initials: Y
authors.lastname: Deng
authors.forename: Liming
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhiyang
authors.initials: Z
authors.lastname: Li
mesh_terms: D001710:Biotin / Q000378:metabolism; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D015894:Genome, Human / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors
chemical_list: D004279:DNA, Viral; D058185:Magnetite Nanoparticles; D001710:Biotin