Amperometric genosensor for culture independent bacterial count.

Bacterial plate count for general assessment of water quality requires lengthy bacterial culturing. We report here a new DNA induced current genosensor for culture independent total bacteria determination. Since the amount of bacterial DNA is correlated to the number of bacteria, the genosensor measures the amount of bacterial DNA to determine bacterial count. The approach relies on bacteria lysis to release DNA which can react with molybdate to form redox molybdophosphate and measured electrochemically. Analysis of E. coli and S. aureus demonstrated that the DNA generated current is highly correlated with the level of bacteria lysis which was confirmed by spectrometric measurement. Culture independent measurement of S. aureus bacterial load suggests limit of detection is 21.9 CFU/mL, with linear range from 3×102 to 3×107 CFU/mL and correlation coefficient of 0.992. For E. coli analysis, the detection limit is 25.1 CFU/mL with the same linear range. The use of electrochemical microbial DNA quantitation for culture independent bacterial count is a new approach, the genosensor measurement is rapid (within 1 h) and has potential use for analysis of broad-spectrum bacteria for various applications.