Enzyme-linked immunoadsorbent assay with electrochemical detection of .alpha.1-acid glycoprotein

Doyle, Matthew; Halsall, H. Brian; Heineman, William R.

doi:10.1021/ac00277a022

Cited by 63 publications

(20 citation statements)

References 64 publications

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“…Enzyme-labels have long been used in electrochemical enzyme-linked immunosorbent assays as a means of amplifying the electrochemical signal for bioaffinity interactions [22]. Similarly, enzyme labels can be applied to electrochemical DNA assays for hybridization detection by labeling the target DNA sequence (simple assay) or reporter DNA probe (sandwich assay) with a redox-active enzyme [23 -40].…”

Section: Introductionmentioning

confidence: 99%

Enzyme‐Amplified Amperometric Detection of DNA Using Redox Mediating Films on Gold Microelectrodes

Hajdukiewicz¹,

Boland²,

Kavanagh³

et al. 2009

Electroanalysis

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Here we describe an amperometric assay for enzyme-labeled detection of DNA hybridization based on a redox polymer film, cross-linked and co-immobilized, with a 20-mer oligonucleotide, to pre-adsorbed cysteamine on gold microelectrode surfaces. Hybridization between the immobilized probe DNA and a biotin-modified target DNA (designed from the ssrA gene of Listeria monocytogenes), followed by addition of an enzyme (glucose oxidase)-avidin conjugate and glucose, results in bioelectrocatalytic oxidation of glucose mediated by the redox polymer, generating significant amplification. The use of gold microelectrodes (25 mm, 40 mm, 100 mm diameter), coupled to careful preparation of surfaces, electronic shielding and background subtraction, results in improved analytical performance, compared to that at macroelectrodes, yielding a limit of quantification of ca. 0.6 pM, corresponding to the presence of ca. 2.5 million copies in the 7 mL assay droplet.

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Section: Introductionmentioning

confidence: 99%

Enzyme‐Amplified Amperometric Detection of DNA Using Redox Mediating Films on Gold Microelectrodes

Hajdukiewicz¹,

Boland²,

Kavanagh³

et al. 2009

Electroanalysis

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“…Phenol is not normally expected to be present in food sampies or microbiological media and this method should, and appears to, suffer little interference from samples. Previous immunoassays employing this principle (Doyle et al 1984;Wehmeyer et al 1985) incorporated column or liquid chromatography steps. In this paper we have been able to successfully apply this method directly to samples and the electrochemical detection step was very simple and rapid (about 2 min/sample).…”

Section: Discussionmentioning

confidence: 99%

A semi-homogeneous amperometric immunosensor for protein A-bearing Staphylococcus aureus in foods

Mirhabibollahi

Brooks

Kroll

1990

Appl Microbiol Biotechnol

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A semi-homogeneous amperometric immunosensor specific to the protein A of Staphylococcus aureus was developed using direct electrochemical detection of phenol produced by alkaline phosphatase from phenyl phosphate. The immunosensor could reliably detect strains of protein A-bearing S. aureus in pure cultures at ca. 10(4) cfu/ml, and at ca. 10(5) cfu/g or ml in various food samples. Due to its semi-homogeneous nature, the system was very simple, easy to operate, and labour-saving. The good correlation between the amperometric current generated by the immunosensor and plate counts illustrated the potential usefulness of this simple system. It proved to be a reliable 24-h detection method for food samples containing very low numbers of protein A-bearing S. aureus after pre-enrichment, as it was able to detect cells that could not directly be enumerated by plate counts.

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“…Enzyme-linked immunosorbent assay (ELISA) is currently one of the most predominant analytical tools for the quantitative determination of a broad variety of analytes in clinical [1][2][3], medical [4], biotechnological [5], and environmental significance [6,7], because of its specificity and sensitivity of antigen-antibody interactions. It is common and routine to perform immunoassay on a microtiter plate with typically 48 or 96 sample wells.…”

Section: Introductionmentioning

confidence: 99%

Microchip-based ELISA strategy for the detection of low-level disease biomarker in serum

Liu

Wang

Huang

et al. 2009

Analytica Chimica Acta

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Enzyme-linked immunoadsorbent assay with electrochemical detection of .alpha.1-acid glycoprotein

Abstract: An enzyme-linked Immunoadsorbent voltammetric assay based on the detection of catalytlcally generated electroactive product has been Investigated. Human orosomucold (OMD,

Cited by 63 publications

References 64 publications

Enzyme‐Amplified Amperometric Detection of DNA Using Redox Mediating Films on Gold Microelectrodes

Enzyme‐Amplified Amperometric Detection of DNA Using Redox Mediating Films on Gold Microelectrodes

A semi-homogeneous amperometric immunosensor for protein A-bearing Staphylococcus aureus in foods

Microchip-based ELISA strategy for the detection of low-level disease biomarker in serum

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