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Label‐Free and Label Based Electrochemical Detection of Hybridization by Using Methylene Blue and Peptide Nucleic Acid Probes at Chitosan Modified Carbon Paste Electrodes
Author(s) -
Kara Pinar,
Kerman Kagan,
Ozkan Dilsat,
Meric Burcu,
Erdem Arzum,
Nielsen Peter E.,
Ozsoz Mehmet
Publication year - 2002
Publication title -
electroanalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 128
eISSN - 1521-4109
pISSN - 1040-0397
DOI - 10.1002/elan.200290011
Subject(s) - peptide nucleic acid , methylene blue , oligonucleotide , dna , nucleic acid , guanine , biosensor , chitosan , combinatorial chemistry , chemistry , hybridization probe , nucleic acid thermodynamics , electrode , carbon paste electrode , oligomer restriction , detection limit , dna–dna hybridization , methylene , electrochemistry , nucleotide , biochemistry , chromatography , cyclic voltammetry , organic chemistry , gene , catalysis , photocatalysis , base sequence
A chitosan modified carbon paste electrode (ChiCPE) based DNA biosensor for the recognition of calf thymus double stranded DNA (dsDNA), single stranded DNA (ssDNA) and hybridization detection between complementary DNA oligonucleotides is presented. DNA and oligonucleotides were electrostatically attached by using chitosan onto CPE. The amino groups of chitosan formed a strong complex with the phosphate backbone of DNA. The immobilized probe could selectively hybridize with the target DNA to form hybrid on the CPE surface. The detection of hybridization was observed by using the label‐free and label based protocols. The oxidation signals of guanine and adenine greatly decreased when a hybrid was formed on the ChiCPE surface. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with target. The signals of MB were investigated at dsDNA modified ChiCPE and ssDNA modified ChiCPE and the increased peak currents were observed, in respect to the order of electrodes. The hybridization of peptide nucleic acid (PNA) probes with the DNA target sequences at ChiCPE was also investigated. Performance characteristics of the sensor were described, along with future prospects.