title: Synthesis of tethered poly(N-isopropylacrylamide) for detection of breast cancer recurrence DNA.
abstract: We have grafted temperature-responsive tethered poly(N-isopropylacrylamide) (PNIPAAm) onto silicon surfaces through atom transfer radical polymerization (ATRP) as a medium to extract human genomic DNA molecules from a biological specimen, namely human blood incorporating target DNA (hgDNA584) and control DNA (hgDNA528) at concentrations of 0.5, 1, and 50 ng μL(-1). The variable adhesion forces of the tethered PNIPAAm brushes on the surfaces were used to capture and release DNA molecules through changes in temperature. After amplifying the signal of the hgDNA584 and hgDNA528 strands released from the tethered PNIPAAm on the substrate using the polymerase chain reaction (PCR), we identified these DNA macromolecules using agarose gel electrophoresis. The accuracy of the detection of hgDNA584 and hgDNA528 was controlled through the design of specific primers in the PCR process. The quantities of these two DNA molecules obtained through the capture and release from tethered PNIPAAm brushes under temperature tuning conditions were sufficient for them to be amplified recognizably, suggesting that this approach could be used in miniaturized lab-on-a-chip cartridges for rapid disease diagnosis.
authors.affiliation: Department of Polymer Engineering, National Taiwan University of Science and Technology, 43, Section 4, Keelung Road, Taipei 106, Taiwan, ROC. jkchen@mail.ntust.edu.tw
authors.forename: Jem-Kun
authors.initials: JK
authors.lastname: Chen
authors.forename: Jun-Yan
authors.initials: JY
authors.lastname: Li
mesh_terms: D000178:Acrylamides / Q000138:chemical synthesis*; D000180:Acrylic Resins; D001943:Breast Neoplasms / Q000175:diagnosis*; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000097:blood* / Q000302:isolation & purification; D005260:Female; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000138:chemical synthesis*; D012008:Recurrence; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D000178:Acrylamides; D000180:Acrylic Resins; D017931:DNA Primers; D004273:DNA, Neoplasm; D011108:Polymers; C052970:poly-N-isopropylacrylamide