Towards an early diagnosis of HIV infection: an electrochemical approach for detection of HIV-1 reverse transcriptase enzyme.
Oriented for the rapid diagnosis of HIV infection, highly sensitive and facile electrochemical assays for HIV-1 reverse transcriptase (HIV1-RT) are presented in this article. A non-labeled and a labeled assay format were based on the formation of self-assembled monolayers (SAMs) of either lipoic acid active ester or a newly synthesized ferrocene (Fc)-labeled cystamine derivative on electrode surfaces, respectively. A short RT-specific peptide, VEAIIRILQQLLFIH, was covalently attached to the surface of the formed SAMs. Electrochemical impedance spectroscopy (EIS) allowed a sensitive interrogation of RT in the non-labeled assay format. Furthermore, square wave voltammetry (SWV) offered a two-dimensional measurement of RT based on the anodic shift and reduction of current density of the Fc redox signal upon binding of RT to its specific peptide. These techniques allowed a linear quantification of the target RT in the range of 75 to 750 pg mL(-1), with a limit of detection of 50 pg mL(-1). Furthermore, the developed biosensors showed a good specificity and allowed a proper discrimination between RT and other HIV enzymes.
The University of Western Ontario, Chemistry Department, London, Canada.
Mahmoud
M
Labib
Patrick O
PO
Shipman
Sanela
S
Martić
Heinz-Bernhard
HB
Kraatz
D015374:Biosensing Techniques / Q000379:methods*; D042241:Early Diagnosis; D004563:Electrochemistry / Q000379:methods*; D015658:HIV Infections / Q000175:diagnosis*; D054303:HIV Reverse Transcriptase / Q000032:analysis*; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D013499:Surface Properties
D054303:HIV Reverse Transcriptase