Visible paper chip immunoassay for rapid determination of bacteria in water distribution system

Paper chips for immunoassay were patterned by screen printing of polydimethylsiloxane (PDMS) or wax pencil drawing. The methods for paper chip patterning are cheap, convenient, rapid and suitable for most laboratories. The whole time for patterning a paper chip is no more than 10 min. Visible immunoassay for the detection of bacteria (Escherichia coli) has been realized using the paper chip, on which the antibody for capturing E. Coli was immobilized on the detection zones of the paper chip, while the detection antibody was labeled with gold nanoparticles (AuNPs) as a signal reporter. After an immunological reaction, the AuNPs bound on the paper chip can effectively catalyse the reduction of silver ions during the silver enhancing step, generating a visible result that can be read by naked eyes. The quantitative results can be acquired by scanning the silver stained paper chip with a commercial scanner/or digital camera. The density of E. coli in water samples can be measured after calibrating the gray value of silver stained spots with the logarithmic number of bacteria. The time and reagents consumed on the paper chip immunoassay is much smaller than those of conventional ELISA, while the sensitivity of the paper chip immunoassay is comparable to conventional ELISA. The technology proposed in this work displays a great potential in the in-situ analysis when daily monitoring of water quality are required. (C) 2013 Elsevier B.V. All rights reserved.