title: DNA electrochemical sensor for detection of PRSS1 point mutation based on restriction endonuclease technique.
abstract: To construct a restriction endonuclease based biosensor technology for PRSS1 genotyping. We designed a thiol-modified hairpin probe where the neck has EcoRI endonuclease recognition sites according to the PRSS1 gene c.410 C>T (p.T137 M) mutation and it was fixed on the gold electrode. Different charge generated by the binding of MB to phosphate groups of DNA before and after hybridization was used for distinguishing the different genotypes and quantity. This showed that the novel sensor can better distinguish the complementary sequence, single-base mismatches, and completely noncomplementary sequences, and the linear range for the logarithm was Y=-0.0242 X+0.1574, R=0.9912(Y=current, X=log target DNA concentration); the detection limit for DNA detection is estimated to be 50 fM.
authors.affiliation: a Department of Laboratory Medicine, First Affiliated Hospital , Fujian Medical University , Fuzhou , China.
authors.forename: Liu
authors.initials: L
authors.lastname: Qicai
authors.forename: Yi
authors.initials: Y
authors.lastname: Qiang
authors.forename: Wu
authors.initials: W
authors.lastname: Wennan
authors.forename: Wang
authors.initials: W
authors.lastname: Yu
authors.forename: Lin
authors.initials: L
authors.lastname: Liqing
authors.forename: Zhao
authors.initials: Z
authors.lastname: Chengfei
authors.forename: Lin
authors.initials: L
authors.lastname: Xinhua
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D015246:Deoxyribonuclease EcoRI / Q000737:chemistry*; D055664:Electrochemical Techniques; D006801:Humans; D057230:Limit of Detection; D017354:Point Mutation; D014357:Trypsin / Q000235:genetics*
chemical_list: D015246:Deoxyribonuclease EcoRI; C489303:PRSS1 protein, human; D014357:Trypsin
keywords: DNA biosensors; digestion; genotyping; hairpin probe