Voltammetric Application of Polypyrrole-Modified Microelectrode Array for the Characterization of DNA Methylation in Glutathione S-Transferase Pi 1.

Direct and efficient label-free voltammetric detection of Glutathione S-Transferase Pi 1 (GSTP1) hypermethylation is reported using a custom developed 16-channel Microelectrode Array chip. The microelectrode array chip is used in a dipstick configuration allowing detection of DNA hybridization in a solution volume of only 0.35 mL. Platinum microelectrode disks (n = 16) 30 µm in diameter have been modified with a polypyrrole bilayer before any contact with the oligonucleotides. The attachment of the 15-mer Probe DNA to the bilayer is random but controlled by the presence of aliphatic tether groups allowing it to form a bidentate complex with the probe DNA. The voltammetric detection procedure of methylated GSTP1-specific target DNA is combined with bisulfite treatment of target DNA. Changes at the interface of the modified microelectrodes in an array configuration are used to record simultaneously cyclic voltammetry on all of the devices. The detection of the hybridization is evaluated statistically for a yes or no event by comparing the changes in recorded cyclic voltammograms before and after exposure to the Target DNA. All cyclic voltammograms of the methylated target show a greater percentage change than those with the non-methylated target exposure and show a greater change in cyclic voltammogram area after methylated target exposure. We observe an average percentage difference of 25.6% ± 4.9 with a variation of 19.1%. These results demonstrate that the fast sensing strategy possesses sensitivity and good specificity. Furthermore, this technology can potentially support rapid, accurate diagnosis and risk assessment of patients with prostate cancer.

Probing for DNA methylation with a voltammetric DNA detector.

A label-free electrochemical detection of DNA hybridization is used for probing synthetic methylated ssDNA 27-mer or 33-mer targets from the GSTP1-gene. The method is based on electrostatic modulation of the anion-exchange kinetics of a polypyrrole bilayer film deposited on platinum-microelectrodes to which a synthetic single-stranded 15-mer GSTP-1 promoter probe DNA has been attached (DNA detector). The effect of the contact of this DNA-detector with non-methylated and methylated complementary DNA sequences in Tris-buffer is compared using cyclic voltammetry (CV). The DNA-hybridization taking place at the electrode surface leads to a significant decrease of the CV area recorded after exposure to complementary target DNA in comparison to the CV change recorded for non-complementary DNA target. The performance of this miniaturized DNA detector was optimized with respect to hybridization time, temperature, and concentration of the target. It was also evaluated with respect to selectivity, sensitivity, and reproducibility. These results are significant for their possible use as a screening test for hypermethylated DNA sequences.

Chemically sensitive field-effect transistor with polyaniline-ionic liquid composite gate.

A sensing layer for a chemically sensitive field-effect transistor (CHEMFET) based on a composite of camphorsulfonic acid (CSA)-doped polyaniline (PANI) and the room-temperature ionic liquid (IL) 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)-imide, BMI(Tf2N), has been developed and characterized for the sensing of ammonia gas. The work function responses of the cast films with and without IL were analyzed by "stepwise" changes of ammonia gas concentration from 0.5 to 694 ppm in air as a function of the mole fraction of IL to PANI. The PANI x CSA/BMI(Tf2N) layers showed enhanced sensitivities, lower detection limits, and shorter response times. There is experimental evidence that PANI forms a charge-transfer complex with imidazolium cation.