A DNA intercalation-based electrochemical method for detection of Chlamydia trachomatis utilizing peroxidase-catalyzed signal amplification.
A sensitive electrochemical DNA detection method for the diagnosis of sexually transmitted disease (STD) caused by Chlamydia trachomatis was developed. The method utilizes a DNA-intercalating agent and a peroxidase promoted enzymatic precipitation reaction and involves the following steps. After hybridization of the target C. trachomatis gene with an immobilized DNA capture probe on a gold electrode surface, the biotin-tagged DNA intercalator (anthraquinone) was inserted into the resulting DNA duplex. Subsequently, the polymeric streptavidin/peroxidase complex was applied to the biotin-decorated electrode. Peroxidase catalyzed 4-chloronaphthol to produce insoluble product, which is precipitated on the electrode surface in the presence of hydrogen peroxide. Cyclic voltammograms with the gold electrode exhibited a peak current of ferrocenemethanol in electrolyte, which decreased in a proportional way to increasing concentration of target DNA owing to insulation of electrode surface by the growing insoluble precipitate. Using this strategy, we were able to detect picomolar concentrations of C. trachomatis gene in a sample taken from a real patient.
Won
Byoung Yeon
BY
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Lee
Dong Woog
DW
Shin
Sung Chul
SC
Cho
Dae-Yeon
DY
Lee
Soo Suk
SS
Yoon
Hyun C
HC
Park
Hyun Gyu
HG
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002692:Chlamydia trachomatis / Q000235:genetics* / Q000302:isolation & purification*; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D004563:Electrochemistry / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007364:Intercalating Agents / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D009195:Peroxidase / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D007364:Intercalating Agents; D004247:DNA; D009195:Peroxidase