Sample Preparation Chip for PCR Detection of Water Borne Pathogens

Enterohemorrhagic Escherichia coli (E. coli O157:H7) is a causative agent of food- and water-borne diseases. Newly developed rapid immunoassay- and PCR-based techniques have been used for detection of E. coli O157:H7 in water and food samples. However, they have proven inadequate, because of the large sample size, an insufficient number of pathogens in the sample, sample splitting, antibody cross reaction, and PCR inhibition caused by sample contaminants. We propose a rapid, sensitive, specific and quantitative test for water-borne pathogens. The method can be employed wherever PCR is used to determine the presence, concentration and/or genetic characteristics of bacterial or viral pathogens. Greatest benefit will be seen in the analysis of samples that are large in relation to the typical 2-5 microliter template volume for PCR, particularly for water quality, food safety, and environment tests. We will develop the method initially to detect E. coli O157:H7 in drinking water in Phase I and source and recreational waters in Phase II. The public will benefit greatly from safer drinking water, rivers and beaches.