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Hybridization assay at a disposable electrochemical biosensor for the attomole detection of amplified human cytomegalovirus DNA.
- Source :
-
Analytical biochemistry [Anal Biochem] 2000 Aug 15; Vol. 284 (1), pp. 107-13. - Publication Year :
- 2000
-
Abstract
- A disposable electrochemical biosensor for the detection of DNA sequences related to the human cytomegalovirus (HCMV) is described. The sensor relies on the adsorption of an amplified human cytomegalovirus DNA strand onto the sensing surface of a screen-printed carbon electrode, and to its hybridization to a complementary single-stranded biotinylated DNA probe. The extent of hybrids formed was determined with streptavidin conjugated to horseradish peroxidase. The peroxidase label was indirectly quantified by measuring the amount of the chromophore and electroactive product 2,2'-diaminoazobenzene generated from the o-phenylenediamine substrate. The intensity of differential pulse voltammetric peak currents resulting from the reduction of the enzyme-generated product was related to the number of target HCMV-amplified DNA molecules present in the sample, and the results were compared to those obtained with standard methods, i.e., agarose gel electrophoresis quantification and colorimetric hybridization assay in a microtiter plate. A detection limit of 0.6 amol/ml of HCMV-amplified DNA fragment was obtained with the electrochemical DNA biosensor. The electrochemical method was 23,000-fold more sensitive than the gel electrophoresis technique and 83-fold more sensitive than the colorimetric hybridization assay in a microtiter plate.<br /> (Copyright 2000 Academic Press.)
- Subjects :
- DNA, Viral blood
Electrophoresis, Agar Gel
Hepacivirus metabolism
Herpesvirus 4, Human metabolism
Humans
Models, Chemical
Polymerase Chain Reaction
Proto-Oncogene Protein c-ets-2
Proto-Oncogene Proteins blood
Spectrophotometry methods
Trans-Activators blood
Biosensing Techniques
Cytomegalovirus genetics
DNA, Viral analysis
DNA-Binding Proteins
Electrochemistry
Nucleic Acid Hybridization methods
Repressor Proteins
Transcription Factors
Subjects
Details
- Language :
- English
- ISSN :
- 0003-2697
- Volume :
- 284
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Analytical biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 10933863
- Full Text :
- https://doi.org/10.1006/abio.2000.4692