A New Biosensor for Rapid Identification of Bacterial Pathogens

EPA Contract Number: 68D02051
Title: A New Biosensor for Rapid Identification of Bacterial Pathogens
Investigators: Tabacco, Mary Beth
Small Business: Echo Technologies Inc.
EPA Contact: Manager, SBIR Program
Phase: II
Project Period: June 1, 2002 through June 1, 2004
Project Amount: $224,965
RFA: Small Business Innovation Research (SBIR) - Phase II (2002) Recipients Lists
Research Category: Ecological Indicators/Assessment/Restoration , SBIR - Monitoring , Small Business Innovation Research (SBIR)

Description:

Rapid, handheld, or portable instrumentation for determining the quality of natural waters, recreational waters, and distributed and treated supplies does not currently exist. Echo Technologies, Inc., completed a Phase I project that demonstrated a new approach for the identification of bacteria in aqueous systems. The approach uses bacteriophage as the molecular recognition element. Bacteriophage are virus particles that generally attach to and infect a narrow range of host cells. Biosensors based on this molecular recognition offer a rapid, selective, and potentially very sensitive method to detect bacteria and bacterial pathogens in potable and recreational waters.

Feasibility of the concept was demonstrated by fabricating fluorescently labeled virus probes (FLVPs) to detect and identify bacteria for representative waterborne bacteria, including: Escherichia coli, Enterococcus faecalis, and Vibrio natriegens bacteria. The preparation of FLVPs has been made quite reproducible. The adhesion to host cells is very rapid, good selectivity was demonstrated in samples with host cells and mixtures, and the response from a solid-phase FLVP biosensor was demonstrated. Several experiments were conducted with a customized detection system to demonstrate the feasibility of making a small in-line instrument capable of high-sensitivity detection. These cumulative results lay an excellent foundation for the Phase II development effort. In Phase II, a prototype rapid bacteria identification system will be designed, fabricated, and evaluated in the laboratory and at an independent test facility. The heart of the instrument will be an array of encoded FLVPs integrated with a charge coupled device (CCD)-based imaging/detection system.

Application of the FLVP technology to solid-state optical sensing represents a new approach to real-time detection of bacterial pathogens. This approach will minimize the need for culturing to identify pathogens, and is an important departure from immunoassay- or DNA-based sensing concepts. The miniature probes are perfectly suited for incorporation in a sensor array for the simultaneous detection of many bacterial pathogens.

Instruments for real-time identification of bacterial pathogens do not currently exist. The FLVP biosensors could be applied to monitoring municipal water supplies, recreational waters, and cooling water towers. They also could benefit both government and civilian users in the area of rapid food safety assessment, particularly in air handling systems, food and beverage processing, and meat processing plants to monitor carcass wash water. The sensors also could be used for monitoring indoor air quality in commercial facilities (e.g., hospitals), as well as in residences.

Supplemental Keywords:

small business, SBIR, water, bacteria, pathogens, fluorescently labeled virus probes, engineering, chemistry, EPA, biosensor, fluorescence spectroscopy, charge coupled device, RFA, INTERNATIONAL COOPERATION, Water, Ecosystem Protection/Environmental Exposure & Risk, POLLUTANTS/TOXICS, Ecosystem/Assessment/Indicators, Ecosystem Protection, Monitoring/Modeling, Ecological Effects - Environmental Exposure & Risk, Drinking Water, Engineering, Chemistry, & Physics, Microorganisms, monitoring, cryptosporidium parvum oocysts, pathogens, microelectromechanical systems (MEMS), biosensing, microbial monitoring, waterborne disease, pathogenic microbes, in situ sensor, cryptosporidium , biosensing system, DNA probe, microorganism, DNA, biosensors, biosensor

Progress and Final Reports:

  • Final Report

  • SBIR Phase I:

    A New Biosensor for Rapid Identification of Bacterial Pathogens  | Final Report