Electrochemical determination of microRNA-21 based on bio bar code and hemin/G-quadruplet DNAenzyme.

A simple and novel microRNA (miRNA) biosensor was developed using DNA-Au bio bar code (DNA-Au) and G-quadruplex-based DNAenzyme. DNA-Au increased the amount of miRNA-21 participating in hybridization. Hemin/G-quadruplex DNAenzyme significantly improved the catalysis of H(2)O(2) by oxidation of hydroquinone, resulting in an obvious reduction current of benzoquinone for miRNA-21 indirect detection. Under optimum conditions, the linear relationship between miRNA-21 concentration and reduction response was obtained with the detection limit of 0.006 pM, which showed a good sensitivity. Besides, selectivity of the biosensor was investigated by detecting the base mismatched miRNAs. This proposed method was further applied to detect miRNA-21 extracted from human hepatocarcinoma BEL-7402 cells and human mastocarcinoma MCF-7 cells. The influence of bisphenol A (BPA) on the expression of miRNA-21 in cells was also investigated. The biosensor performs well in practical applications, which suggests it may provide a new platform for gene diagnosis.

Electrochemical detection of DNA damage induced by acrylamide and its metabolite at the graphene-ionic liquid-Nafion modified pyrolytic graphite electrode.

A new electrochemical biosensor for directly detecting DNA damage induced by acrylamide (AA) and its metabolite was presented in this work. The graphene-ionic liquid-Nafion modified pyrolytic graphite electrode (PGE) was prepared, and then horseradish peroxidase (HRP) and natural double-stranded DNA were alternately assembled on the modified electrode by the layer-by-layer method. The PGE/graphene-ionic liquid-Nafion and the construction of the (HRP/DNA)(n) film were characterized by electrochemical impedance spectroscopy. With the guanine signal in DNA as an indicator, the damage of DNA was detected by differential pulse voltammetry after PGE/graphene-ionic liquid-Nafion/(HRP/DNA)(n) was incubated in AA solution or AA+H(2)O(2) solution at 37°C. This method provides a new model to mimic and directly detect DNA damage induced by chemical pollutants and their metabolites in vitro. The results indicated that, in the presence of H(2)O(2), HRP was activated and catalyzed the transformation of AA to glycidamide, which could form DNA adducts and induce more serious damage of DNA than AA. In order to further verify these results, UV-vis spectrophotometry was also used to investigate DNA damage induced by AA and its metabolites in solution and the similar results were obtained.

Electrochemical detection of DNA hybridization using a change in flexibility.

A novel electrochemical method of detecting DNA hybridization is presented based on the change in flexibility between the single and double stranded DNA. A recognition surface based on gold nanoparticles (GNPs) is firstly modified via mixing self-assembled monolayer of thiolated probe DNA and 1,6-hexanedithiol. The hybridization and electrochemical detection are performed on the surface of probe-modified GNPs and electrode, respectively. Here in our method the charge transfer resistance (R(ct)) signal is enhanced by blocking the surface of electrode with DNA covered GNPs. The GNPs will be able to adsorb on the gold electrode when covered with flexible single stranded DNA (ssDNA). On the contrary, it will be repelled from the electrode, when covered with stiff double stranded DNA (dsDNA). Therefore, different R(ct) signals are observed before and after hybridization. The hybridization events are monitored by electrochemical impedance spectroscopy (EIS) measurement based on the R(ct) signals without any external labels. This method provides an alternative route for expanding the range of detection methods available for DNA hybridization.

Electrochemical behavior and detection of plant hormone 6-benzyl adenine in acetate buffer at mercury electrode.

The electrochemical behavior of 6-benzyl adenine (6-BA) has been studied in 0.1 mol L(-1) HAc-NaAc solution (pH 3.8). Cyclic voltammetry, single-sweep polarography and direct current polarography were employed to clarify the mechanism of the electrode process; the kinetic parameters of the rate-determining step were determined. Reduction of 6-BA involves two pH-dependent processes, corresponding to the overall irreversible reduction of the 1,6 and 3,2 N=C bonds. Each reduction stage consists a preprotonation of the nitrogen atom at the electroactive site and a rapid two-electron transfer. In the presence of 6-BA, the reduction potentials of some ions were shifted. Under the given conditions, 6-BA displays one irreversible reduction peak controlled by adsorption. Two linear response were observed in the range 2.0 x 10(-8) - 8.0 x 10(-7) mol L(-1) and the range 1.0 x 10(-6) - 8.0 x 10(-6) mol L(-1), with correlation coefficients of 0.9995 and 0.9998, respectively. The detection limit is 7.1 x 10(-9) mol L(-1). The method had been applied to the determination of 6-BA in bean sprout samples with satisfactory results.

Fluorometric method for quantitative determination of glucose and its application to human serum.

By coupling the Fenton reaction with the glucose oxidase (GOD)-catalyzed reaction, we have developed a fluorometric method for the determination of glucose; the linear response range and the detection limit are 0.108-2.59 microg/ml and 0.0432 microg/ml, respectively. This approach is selective and has the advantages of using inexpensive reagents and avoiding interferences. Satisfactory results were obtained for the determination of glucose in human serum samples.