title: Conditions of polymerase chain reaction amplification by magnetic enrichment and nanoscale detection sensitivity.
abstract: In this research, we reported a method of polymerase chain reaction (PCR) amplification by means of magnetic enrichment. First, after denaturation, the target sequence was combined with biotin-modified specific primer through hybridization and enriched at the surface of gamma-Fe2O3 by biotin-avidin special interaction. Then single target sequence was gained through denaturation, and general PCR amplification was performed. The experiment conditions such as the hybridization temperature between target sequence and biotin-modified specific primer, and the dosage of magnetic nanoparticles gamma-Fe2O3 were optimized. Finally, the sensitivity of the method was checked. The lowest concentration of target sequence was detected as low as 5 x 10(-7) ng/mL. This simple method could provide a quick and early diagnosis of malignant infectious diseases such as SARS, avian flu and swine flu etc., that occur occasionally nowadays.
authors.affiliation: Hunan Key Laboratory of Green Packing and Application of Biological Nanotechnology, Hunan University of Technology, Zhuzhou 412007, China.
authors.forename: Nongyue
authors.initials: N
authors.lastname: He
authors.forename: Xianbo
authors.initials: X
authors.lastname: Mou
authors.forename: Yan
authors.initials: Y
authors.lastname: Deng
authors.forename: Hongna
authors.initials: H
authors.lastname: Liu
authors.forename: Song
authors.initials: S
authors.lastname: Li
authors.forename: Lijian
authors.initials: L
authors.lastname: Xu
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Li
mesh_terms: D001710:Biotin / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D005290:Ferric Compounds / Q000737:chemistry*; D006801:Humans; D008280:Magnetics / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D005290:Ferric Compounds; D001710:Biotin; D004247:DNA