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Soil and Water – What is Detectable through Microbiological Sample Preparation Techniques
Citation:
KEELEY, ANN AND Q. Wu. Soil and Water – What is Detectable through Microbiological Sample Preparation Techniques. U.S. Environmental Protection Agency, Washington, DC, EPA/600/F-10/021, 2010.
Impact/Purpose:
Objectives are focused on the importance of sample preparation in the rapid, precise, and sensitive identification of biological agents in responding to incidences potentially resulting in civilian harm. Difficulties involved result in large measure because existing methods were developed for clinical sampling.
Description:
The concerns of a potential terrorist’s use of biological agents in soil and ground water are articulated by comparisons to major illnesses in this Country involving contaminated drinking water sources. Objectives are focused on the importance of sample preparation in the rapid, precise, and sensitive identification of biological agents in responding to incidences potentially resulting in civilian harm. Difficulties involved result in large measure because existing methods were developed for clinical sampling. The importance of sample collection, transport media, and matrix preparation are highlighted along with the validation of analyte-specific filtration, purification, and staining procedures. Both molecular and culture-based techniques are discussed in accurately demonstrating when soil and water samples harbor potential pathogens. Salmonella enterica Serovars Typhi, Cryptosporidium parvum, and Giardia lamblia were selected for these investigations using water and soil samples. Real time PCR was used to replace traditional PCR as it is more rapid, precise and more sensitive. Efforts to design and test primers/probes is discussed along with those to find optimal conditions of testing. Detection limits are discussed with respect to analytical procedures and future activities are highlighted.