Determination of nanogram quantities of osmium-labeled nucleic acids by stripping (inverse) voltammetry

Paleček, Emil; Hung, Mac Anh

doi:10.1016/0003-2697(83)90002-7

Cited by 79 publications

(48 citation statements)

References 8 publications

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“…In electrochemical DNA hybridization sensors a covalently bound electroactive label is frequently introduced in the reporter (signaling) probe (RP) [1 -3], in target DNA (tDNA) [4] or in the capture probe [5 -7]. To our knowledge, the first electroactive markers of this type were osmium tetroxide complexes, which were introduced in DNA and RNA already at the beginning of the 1980s [8,9].…”

Section: Introductionmentioning

confidence: 99%

“…For detection of DNA modified with Os,L at mercury electrodes, the well developed signal due to catalytic evolution of hydrogen is used; its current response is many times higher than those of other steps controlled by rate of diffusion [8,9,19]. A considerable increase of sensitivity of analyses (up to several orders of magnitude) can be achieved by the application of stripping voltammetry where detection limit of denatured, osmium tetroxide-modified calf thymus DNA of 5 ng mL À1 was reached [9]. The adsorption of NA on surface of the detection (working) electrode is sufficiently strong to allow preparation of a DNA-modified electrode resisting washing [16,22,23].…”

Section: Introductionmentioning

confidence: 99%

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Voltammetric Behavior of Osmium-Labeled DNA at Mercury Meniscus-Modified Solid Amalgam Electrodes. Detecting DNA Hybridization

et al. 2006

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The complex of osmium tetroxide with 2,2'-bipyridine has been utilized as a probe of DNA structure and an electroactive marker of DNA in DNA hybridization sensors. It produces several voltammetric signals, the most negative of them has been observed only at mercury electrodes. This signal is of catalytic nature affording a high sensitivity of DNA determination. The catalytic current due to evolution of hydrogen in voltammetry of DNA modified by complex of osmium tetroxide with 2,2'-bipyridine (DNA-Os,bipy) was studied. Solid amalgam electrodes (modified with mercury menisci) of silver (m-AgSAE), copper (m-CuSAE), gold, and of combined bismuth and silver, were used as possible substitutes for mercury electrodes. Besides the hanging mercury drop electrode (HMDE), the catalytic current was observed only on m-AgSAE and m-CuSAE. Electrodes of gold and bismuth amalgams did not give the catalytic current. The detection limit of DNA-Os,bipy on HMDE was 0.1 ng mL À1 (RSD ¼ 2.3 %, N ¼ 11), and on m-AgSAE 0.2 ng mL À1 (RSD ¼ 3.1%, N ¼ 11). The m-AgSAE was successfully applied as a detection electrode in double-surface DNA hybridization experiments offering highly specific discrimination between complementary (target) and nonspecific DNAs, as well as determination of the length of a repetitive DNA sequence. The m-AgSAE has proved a convenient alternative to the HMDE or carbon electrodes used for similar purposes in previous work.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Voltammetric Behavior of Osmium-Labeled DNA at Mercury Meniscus-Modified Solid Amalgam Electrodes. Detecting DNA Hybridization

et al. 2006

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“…Osmium tetroxide complexes (Os,L) were introduced as electroactive DNA markers in the early 80×s [18,19].…”

Section: Introductionmentioning

confidence: 99%

“…This feature makes Os, bipy and some other Os,L excellent DNA structural probes [20,23]. At the mercury or carbon electrodes, Os,L as well as Os,L-modified DNA (DNA-Os,L) undergo several (quasi)reversible faradaic processes corresponding to consecutive reduction/oxidation of osmium atom [19,21,22,24]. We have recently shown [22] that DNA-Os, bipy yields a specific peak (peak a) at the pyrolytic graphite electrode (PGE) whose potential significantly differs from that of a corresponding signal of free Os, bipy reagent.…”

Section: Introductionmentioning

confidence: 99%

“…This procedure was ineffective in connection with the mercury electrode; measurements of osmium-labeled DNA at the HMDE required careful removal of the free Os, bipy reagent prior to analysis. On the other hand, measurement of a catalytic osmium peak yielded by the modified DNA at HMDE represents probably the most sensitive DNA electrochemical determination without previous degradation (down to sub-femtomole amounts when 5-microliter samples are analyzed) [19,24].…”

Section: Introductionmentioning

confidence: 99%

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Two‐Surface Strategy in Electrochemical DNA Hybridization Assays: Detection of Osmium‐Labeled Target DNA at Carbon Electrodes

et al. 2003

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Target DNAs, including a 71-mer oligonucleotide, a PCR product and a plasmid DNA, all containing oligo(A) stretches, were hybridized at magnetic Dynabeads oligo(dT) 25 (DBT). The hybridization events were detected using a technique based on chemical modification of the target DNA with a complex of osmium tetroxide with 2,2'-bipyridine (Os, bipy) and voltammetric detection at carbon electrodes. DNA was modified with Os, bipy prior to capture at DBT, at the beads, or after release from the beads. In the latter case, DNA-Os, bipy was detected in the reaction mixture using adsorptive transfer stripping voltammetry involving extraction of unreacted Os, bipy from the electrode by organic solvents. Pre-labeling of the target plasmid DNA and the PCR product with Os, bipy significantly increased the yield of DNA captured at the beads. Tens of femtomoles of both short (the 71-mer oligonucleotide) and long (the 3-kilobase plasmid) target DNAs in a 20-microliter hybridization sample can be easily detected by means of these techniques. Various carbon electrode materials, including pyrolytic graphite (PGE), highly oriented pyrolytic graphite (HOPGE), carbon paste (CPE), glassy carbon and pencil graphite, were tested regarding their suitability for the detection of osmium-labeled DNA. Among them, PGE and HOPGE appeared usable in the measurements of both purified DNA-Os, bipy and its mixtures with unreacted Os, bipy while CPE was suitable for the detection purified osmium-labeled DNA.

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Electrochemical Sensors for DNA Interactions and Damage

2002

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Determination of nanogram quantities of osmium-labeled nucleic acids by stripping (inverse) voltammetry

Cited by 79 publications

References 8 publications

Voltammetric Behavior of Osmium-Labeled DNA at Mercury Meniscus-Modified Solid Amalgam Electrodes. Detecting DNA Hybridization

Voltammetric Behavior of Osmium-Labeled DNA at Mercury Meniscus-Modified Solid Amalgam Electrodes. Detecting DNA Hybridization

Two‐Surface Strategy in Electrochemical DNA Hybridization Assays: Detection of Osmium‐Labeled Target DNA at Carbon Electrodes

Electrochemical Sensors for DNA Interactions and Damage

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