title: Highly sensitive and multiple DNA biosensor based on isothermal strand-displacement polymerase reaction and functionalized magnetic microparticles.
abstract: A universal, highly sensitive and selective chemiluminescence (CL) imaging method has been developed for high throughput detection of DNA. After molecular beacon (MB) hybridized with the target DNA, the biotin-labeled primer was attached to a magnetic microparticle (MMP) surface by hybridization with the stem part of the MB to initiate a polymerization of DNA strand, which led to the release of the target and another polymerization cycle. Thus the polymerization produced the multiplication of biotin-labeled primer on the surface of MMPs. Sequentially, the horseradish peroxidase (HRP) was conjugated to MMPs surface through the biotin-streptavidin reaction. Then, the conjugated HRP was determined by the CL imaging method. This proposed method could detect the sequence-specific DNA as low as 0.4 pM and discriminate perfectly matched target DNA from the mismatch DNAs. All in all, this proposed method exhibited an efficient amplification performance, and would open new opportunities for sensitive and high throughput detection of DNA.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Luo
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Ningxing
authors.initials: N
authors.lastname: Li
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Yufei
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Chaohui
authors.initials: C
authors.lastname: Chen
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Xia
authors.initials: X
authors.lastname: Xiang
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China. Electronic address: zhkhe@whu.edu.cn.
authors.forename: Xinghu
authors.initials: X
authors.lastname: Ji
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Zhike
authors.initials: Z
authors.lastname: He
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation*; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017403:In Situ Hybridization / Q000295:instrumentation*; D008163:Luminescent Measurements / Q000295:instrumentation*; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D058185:Magnetite Nanoparticles; D004247:DNA
keywords: CL imaging; DNA biosensors; Polymerase; Signal amplification