Progress 05/15/03 to 11/14/04
Outputs
Biological warfare (bioterrorism) agents (BWAs) are an ever-increasing threat. Homeland security has become one of the U.S.A.'s top priorities following recent terrorist activities. The CDC has identified several agents that are potential bioterrorism threats. To ensure homeland security, new technologies and tools are needed that will help rural communities become more resilient when faced with bioterrorism related disasters. Rural communities, due to their remote locations, have the minimal infrastructure and capability to counteract terrorist attacks, so the need to have methods for dealing with disasters is critical. Due to the urgent needs for new technology, Maxwell Sensors Inc. (MSI) proposed to develop a miniaturized enzyme linked immunosorbent assay (ELISA) based biochip and platform, that can provide rapid, & simultaneous detection and identification of prevalent bioterrorism and agriculture related pathogens with ultra high sensitivity and accuracy. ELISA is
widely used for laboratory confirmatory tests. ELISA is a very sensitive and specific method, however it is labor intensive, and requires both large systems and highly trained personnel to operate them. The uniqueness of MSI's ELISA-chip is that it combines: 1. The 'gold standard' ELISA method as an assay and detection principle. 2. MSI's proprietary self-contained microfluidic chip technology in order to automate the assay process and miniaturize the system for rural community health care applications. This ELISA system utilizes microfabrication and integrated microfluidics to provide biochemistry automation. The ELISA chip with, pre-loaded reagents, is not only ready for use and convenient for rapid assays, but also eliminates the need for a network of tubing connected to external reagent reservoirs and bulky pump systems. The system utilizes a microactuator to release reagents from sealed compartments, and control both fluid dynamics and biochemical reactions, thus reducing the
operational procedure to a single step. During Phase I of the project, we have: 1. Developed the ELISA based assay protocol for BWA tests 2. Designed and fabricated the ELISA based microfluidic platform 3. Constructed the pressure driven micro mechanism for total automation 4. Integrated the BWA-Chip system in order to provide rapid testing (< 25 minutes) 5. Performed bioassays and characterized the system with: * Bacillus globigii (BG): Bacillus Anthracis simulant & achieved excellent sensitivity (< 56 organisms or < 0.1ng) (Sec. 3.2.4.1) * Staphylococcal enterotoxin B (SEB): (Sec. Sec. 3.2.4.2) * E. Coli O157:H7 food Safety agent (Sec. Sec. 3.2.4.3) * Salmonella food safety agent (Sec. Sec. 3.2.4.4) 6. Evaluated the technical merits and commercial potential 7. Successfully demonstrated the platform's technical feasibility.
Impacts
Currently, there are three urgent problems: 1. The infective dose of many BWAs is quite low and the incubation period is often short. Unfortunately current clinical diagnostic methods require highly trained personnel and take 3 to 6 days to produce results. 2. Even low levels of BWA can pose a contamination threat to people. Rapid environmental sampling (water, food, air, etc.) in order to ascertain the presence, or absence, of bioterrorism related agents, is critical for assessing the risk of exposure. Unfortunately technologies for rapidly assessing pathogens in the environment remain underdeveloped. 3. Consumers have been worried about food safety and agriculture related diseases. Food borne pathogens (e.g. salmonella, E coli 0157:H7) and zoonotic diseases (e.g. hoof and mouth disease) have the potential to cause illness or death. During Phase I of the project, we developed a microfluidic based ELISA chip for sensitive biological agent assay, investigated & selected
immuno-specific agents and materials, constructed a laboratory BWA system, characterized the system with direct detection of biological agents (e.g. bacillus anthracis stimulant- Bacillus globigii (BG) spore, Staphylococcal enterotoxin B (SEB), E. coli O157:H7, and Salmonella), performed control sample tests, and demonstrated the multiplexing assay capability by processing six samples simultaneously. In conclusion, the proposed ELISA Chip is lightweight, compact, inexpensive, and can provide both 'rapid detection' and 'sensitivity'.
Publications
- No publications reported this period
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