title: Nano-magnetic primer based electrochemiluminescence-polymerase chain reaction (NMPE-PCR) assay.
abstract: Here we have developed a novel nano-magnetic primer based electrochemiluminescence-polymerase chain reaction (NMPE-PCR) strategy for detection of genome. The key idea of this method is integrating the two in situ processes: PCR on the surface of magnetic nanoparticles (MNPs) and magnetic beads based ECL readout platform, to avoid some laborious manual operations and achieve rapid yet sensitive detection. At first, the approach employs a pair of functional primers for amplification: one is tris-(2,2'-bipyridyl) ruthenium (TBR) labeled primer; the other one is nano-magnetic primer which is prepared by attaching the primer to the surfaces of MNPs. With the presence of DNA analyte and PCR mixture, the TBR labeled products are directly loaded and enriched on the surface of MNPs during PCR cycling. Then the MNPs-TBR complexes can be analyzed by a magnetic ECL platform without any post-modification or post-incubation. Finally, we used Listeria monocytogenes as the target to examine these desirable properties of this assay, reaching a detection limit of 500 fg/μL for genome in 1 h. The proposed study has provided the evidence as a proof-of-concept, thus having potential for development of automatic mode for detection of specific gene.
authors.affiliation: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhu
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Zhou
authors.forename: Da
authors.initials: D
authors.lastname: Xing
mesh_terms: D002874:Chromosome Mapping / Q000295:instrumentation; D003217:Conductometry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D016680:Genome, Bacterial / Q000235:genetics*; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D008089:Listeria monocytogenes / Q000235:genetics*; D008163:Luminescent Measurements / Q000295:instrumentation*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000295:instrumentation; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D058185:Magnetite Nanoparticles