Biotechnology Reports (Dec 2020)
Rapid detection of mecA gene of methicillin-resistant Staphylococcus aureus by a novel, label-free real-time capacitive biosensor
Abstract
This work presents a rapid, selective and sensitive automated sequential injection flow system with a capacitive biosensor for detection of the mecA gene (the model chosen for this study), which emerges from methicillin-resistant Staphylococcus aureus. A DNA-based 25-mer capture probe was immobilized on the surface of a gold electrode which was integrated in the capacitive sensor system. A constant current pulse was applied and the resulting capacitance was measured. Injection of the target DNA sample to the sensor surface induced hybridization to occur between the target and the complementary sequence, which resulted in a shift in the measured capacitance (ΔC). The ΔC was directly proportional to the concentrations of the applied target probe with linearity ranging from 10−12 to 10−7 M. The biosensor had a detection limit of 6.0 × 10−13 M and a recovery of 95 % of the mecA gene when spiked in human saliva. The biosensor showed a promising selectivity. It could clearly discriminate single-base, two-base and twelve-base mismatch probes with a decrease in the signal strength by 13 %, 26 %, and 89 %, respectively relative to the signal strength of the complementary target probe. There was no significant signal observed for the non-complementary probe. The biosensor-chip could be re-used for more than 12 cycles with residual capacity of 94.5 ± 4.3 % and a RSD of 4.6 % by regenerating the biosensor-chip with a solution of 50 mM NaOH.
