Electrochemical probe DNA design in PCR amplicon sequence for the optimum detection of microbiological diseases

Kara, Pınar; Çavdar, Safiye; Meric, Buket; Erensoy, Selda; Ozsoz, Mehmet

doi:10.1016/j.bioelechem.2007.05.001

Cited by 31 publications

(16 citation statements)

References 29 publications

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“…After hybridization, the biosensor were immersed in washing buffer containing 2xSSC+0.1 % SDS (30 mM sodium citrate, 0.3 M NaCl, 0.1 % sodium dodecyl sulfate, pH 7.0) for 5 min to keep from the unspecific bounding [24]. Since the highest ratio of complementary signal to Fig S1a).…”

Section: Hybridizationmentioning

confidence: 99%

A Nucleic Acid Biosensor for Detection of Hepatitis C Virus Genotype 1a Using Poly(l-Glutamic Acid)-Modified Electrode

Dönmez

Arslan

2015

Appl Biochem Biotechnol

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An electrochemical nucleic acid biosensor based on label-free DNA detection method was prepared for the first time by using electropolymerized poly(L-glutamic acid)-modified pencil graphite electrode (PGA/PGE) for detection of hepatitis C virus genotype 1a (HCV1a). Inosine-substituted 20-mer probes related to the HCV1a were immobilized onto PGA/PGE surface by covalent linking with the formation of amide bonds. Square wave voltammetry (SWV) was used to monitor the oxidation signal of guanine in the hybridization events, which gave an oxidation peak at +1.05 V. An increase in the oxidation signal of guanine was showed by hybridization of the probe with the complementary DNA. Noncomplementary oligonucleotides were also used to investigate the selectivity of the biosensor. The proposed nucleic acid biosensor was linear in the range of 50 nM to 1.0 μM, exhibiting a limit of detection of 40.6 nM. Finally, single-stranded synthetic PCR product analogues of HCV1a were performed in optimal condition. This PGA-modified nucleic acid sensor is cost-effective and disposable, and besides, it has superior electrocatalytic effect on the oxidation of guanine.

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Section: Hybridizationmentioning

confidence: 99%

A Nucleic Acid Biosensor for Detection of Hepatitis C Virus Genotype 1a Using Poly(l-Glutamic Acid)-Modified Electrode

Dönmez

Arslan

2015

Appl Biochem Biotechnol

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“…Th e probe oligonucleotides, having the 5 ′ NH 2 (CH 2 ) 6 link, were immobilized onto the PGE surface by covalent linking. Immobilization was carried out through the formation of amide bonds between the carboxylic functionality at PGE surfaces and the amino-terminal ends of the probe oligonucleotides (Kara et al 2007). Hybridization Th e nucleic acid biosensor was immersed into the hybridization solution (2xSSC solution of pH 7.0 containing the target or non-complementary sequences) for the desired time.…”

Section: Probe Immobilizationmentioning

confidence: 99%

Sequence-specific label-free nucleic acid biosensor for the detection of the hepatitis C virus genotype 1a using a disposable pencil graphite electrode

Dönmez

Arslan

2015

Artificial Cells, Nanomedicine, and Biotechnology

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In this paper, we demonstrate a simple, sensitive, inexpensive, disposable and label-free electrochemical nucleic acid biosensor for the detection of the hepatitis C virus genotype 1a (HCV1a). The nucleic acid biosensor was designed with the amino-linked inosine-substituted 20-mer probes, which were immobilized onto a disposable pencil graphite electrode (PGE) by covalent linking. The proposed nucleic acid biosensor was linear in the range of 0.05 and 0.75 μM, exhibiting a limit of detection of 54.9 nM. The single-stranded synthetic PCR product analogs of HCV1a were also detected with satisfactory results under optimal conditions, showing the potential application of this biosensor.

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“…They were also able to detect the lamivudine resistance, which could provide fast and cost-efficient point-of-care testing in hospitals. Kara et al [40] used PGEs with the capture probe electrostatically and chemically immobilized on it, studying sequences from the HBV genome as target DNA. The influence of the probe design for the optimum detection of diseases caused by HBV was investigated, and it was found that the best performance occurred when the probe was homologous to the middle of the amplicon.…”

Section: Real Samplesmentioning

confidence: 99%

Sequence-specific electrochemical detection of nucleic acids in real samples

Duwensee¹,

Mix

Flechsig

2010

Bioanal Rev

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This paper reviews past and current developments in the field of electrochemical biosensors with a focus on the sequence-specific detection of nucleic acids in real samples. After electrochemical hybridization sensors had been first described in 1993, it took nearly a decade until some of the many proposed protocols were indeed applied to real samples like blood or tissue. Electrochemical transduction schemes used either rely on electroactive moieties such as intercalators, groove binders, covalently attached labels, and products of enzyme markers or they are completely indicator free like impedance-based detection principles. Most detection schemes require a polymerase chain reaction amplification step to allow for sufficient selectivity and sensitivity. Today, several companies develop electrochemical microarrays able to detect dozens to many thousands of sequences in a single experiment.

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Electrochemical probe DNA design in PCR amplicon sequence for the optimum detection of microbiological diseases

Cited by 31 publications

References 29 publications

A Nucleic Acid Biosensor for Detection of Hepatitis C Virus Genotype 1a Using Poly(l-Glutamic Acid)-Modified Electrode

A Nucleic Acid Biosensor for Detection of Hepatitis C Virus Genotype 1a Using Poly(l-Glutamic Acid)-Modified Electrode

Sequence-specific label-free nucleic acid biosensor for the detection of the hepatitis C virus genotype 1a using a disposable pencil graphite electrode

Sequence-specific electrochemical detection of nucleic acids in real samples

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