2000 Progress Report: A New Microfluidic System for the Determination of Cryptosporidium Oocysts in WaterEPA Contract Number: 68D01064
Title: A New Microfluidic System for the Determination of Cryptosporidium Oocysts in Water
Investigators: Hodko, Dalibor
Current Investigators: Hitchens, G. Duncan
Small Business: Lynntech Inc.
EPA Contact: Richards, April
Project Period: September 1, 2001 through September 1, 2003
Project Period Covered by this Report: September 1, 1999 through September 1, 2000
Project Amount: $225,000
RFA: Small Business Innovation Research (SBIR) - Phase II (2001) Recipients Lists
Research Category: Ecological Indicators/Assessment/Restoration , SBIR - Monitoring , Small Business Innovation Research (SBIR)
Description:Waterborne infectious disease outbreaks have been attributed to a variety of pathogenic bacteria, parasites, and viruses. Cryptosporidium outbreaks have been reported less frequently, but the number of cases associated with an outbreak has been much larger. Such outbreaks prompted the U.S. Environmental Protection Agency (EPA) to establish the Information Collection Rule (ICR), which requires that all large public water systems routinely conduct microbiological monitoring of raw and finished waters. Currently available methods for detection of Cryptosporidium oocyst and Giardia cysts, the ICR method and Method 1622, yield low oocysts and/or cysts recovery, cannot be performed in the field, do not distinguish between live and dead protozoa, and need highly trained professionals to accurately perform the analysis. The accuracy of these fluorescent microscopically based methods is strongly affected by nonspecific fluorescence, other organisms, the presence of a chlorine compound, and other physical conditions of the sample. The new method proposed is based on two innovations, one for the separation of live Cryptosporidium oocysts from the water sample and the other for the detection of Cryptosporidium DNA amplicons using a new detection method of polymerase chain reaction (PCR) amplified DNA. The system involves the development of a microfluidic analyzer that will allow highly specific and sensitive detection of Cryptosporidium and other microogranisms and performance of the analysis in the field. The main features of the analyzer include direct interfacing between large sample volume and a microliter volume used in the microfluidic detection channel, on-chip cell lysis, amplification of DNA through rapid thermal cycling, and nonfluorescence on-ship detection of DNA amplicons. A fully developed system should provide unparalleled sensitivity and selectivity for detecting Cryptosporidium oocyst and will not require highly trained personnel for its performance in the field.
The proposed microfluidic system will find a large number of applications in future microfluidics-based instrumentation for integration of basic components for concentration, transport, and detection of biologic and inorganic particles in liquid samples. Potential commercial applications of the developed instrument will include monitoring of microorganisms in the field or on production lines in the food, chemical, pharmaceutical, and biotechnology industries.