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Cited by 273

The DOI matches its Crossref record; 19 of the 19 references it cites were found in a registry or an index.

Electrochemical Genosensor Based on Colloidal Gold Nanoparticles for the Detection of Factor V Leiden Mutation Using Disposable Pencil Graphite Electrodes

Ozsoz, Mehmet, Erdem, Arzum, Kerman, Kagan, Ozkan, Dilsat, Tugrul, Berrin, Topcuoglu, Nejat, Ekren, Hayati, Taylan, Muzaffer·2003·Analytical Chemistry
DOI 10.1021/ac026212r
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Abstract

Electrochemical genosensors for the detection of the Factor V Leiden mutation from polymerase chain reaction (PCR) amplicons using the oxidation signal of colloidal gold (Au) is described. A pencil graphite electrode (PGE) modified with target DNA, when hybridized with complementary probes conjugated to Au nanoparticles, responded with the appearance of a Au oxide wave at approximately +1.20 V. Specific probes were immobilized onto the Au nanoparticles in two different modes: (a) Inosine-substituted probes were covalently attached from their amino groups at the 5' end using N-(3-dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent onto a carboxylate-terminated l-cysteine self-assembled monolayer (SAM) preformed on the Au nanoparticles, and (b) probes with a hexanethiol group at their 5' phosphate end formed a SAM on Au nanoparticles. The genosensor relies on the hybridization of the probes with their complementary targets, which are covalently immobilized at the PGE surface. Au-tagged 23-mer capture probes were challenged with the synthetic 23-mer target, 131-base single-stranded DNA or denatured 256-base polymerase chain reaction (PCR) amplicon. The appearance of the Au oxidation signal shortened the assay time and simplified the detection of the Factor V Leiden mutation from PCR amplified real samples. The discrimination between the homozygous and heterozygous mutations was also established by comparing the peak currents of the Au signals. Numerous factors affecting the hybridization and nonspecific binding events were optimized. The detection limit for the PCR amplicons was found to be as low as 0.78 fmol; thus, it is suitable for point-of-care applications.

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Metadata

Authors
Ozsoz, M., Erdem, A., Kerman, K., Ozkan, D., Tugrul, B., Topcuoglu, N., Ekren, H., & Taylan, M.
Venue
Analytical Chemistry
Publisher
American Chemical Society (ACS)
Published
2003-04-03
Volume
75
Issue
9
Pages
2181-2187
ISSN
  • 0003-2700 (ISSN-L)
  • 1520-6882
DOI
10.1021/ac026212r
OpenAlex
W2048769537
Cited by
273
References
19
FWCI
6.33
Type
journal article
Language
en
Open access
closed

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