title: Detection and discrimination of recombinant plasmid encoding hepatitis C virus core/E1 gene based on PNA and double-stranded DNA hybridization.
abstract: Development of an electrochemical DNA biosensor for direct detection and discrimination of double-stranded plasmid (ds-Pl) without the need for denaturation of the target plasmid sample using a peptide nucleic acid (PNA) oligomer as the probe is described. This goal was achieved by modification of gold electrode with 6-mercapto-1-hexanol following monolayer self-assembly of cysteine conjugated 20-mer PNA oligomer probe, complementary to the HCV core/E1 region, which binds to ds-Pl and forms PNA/ds-Pl structure. The significant variation in differential pulse voltammetric response of methylene blue on the probe modified electrode upon contacting with complementary double-strand plasmid to form PNA/ds-Pl triplex structure is the principle of target plasmid detection. The results indicated that the reduction peak current was linear with the concentration of complementary strand in the range of 10-300 pg/μl with a detection limit of 9.5 pg/μl.
authors.affiliation: Electroanalytical Chemistry Laboratory, Faculty of Chemistry, University of Tabriz, Tabriz, Iran. fatemeh.ahour@gmail.com
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Ahour
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Pournaghi-Azar
authors.forename: Esmaeel
authors.initials: E
authors.lastname: Alipour
authors.forename: Mohammad Saeid
authors.initials: MS
authors.lastname: Hejazi
mesh_terms: D015374:Biosensing Techniques; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004563:Electrochemistry / Q000379:methods; D006046:Gold / Q000737:chemistry; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification; D006526:Hepatitis C / Q000235:genetics* / Q000821:virology; D006801:Humans; D057230:Limit of Detection; D009693:Nucleic Acid Hybridization; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D020135:Peptide Nucleic Acids; D014764:Viral Proteins; D006046:Gold; D004247:DNA