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ELISA MEASUREMENT FOR STACHYLYSIN
Citation:
Van Emon, J M. AND S J. Vesper. ELISA MEASUREMENT FOR STACHYLYSIN. Presented at American Society for Testing Materials, Boulder, CO, July 25-30, 2004.
Impact/Purpose:
More cost-effective field screening and monitoring methods will be provided to increase the amount of information available concerning the location, source, and concentration of pollutants. Rapid and sensitive immunoassays such as enzyme-linked immunosorbent assays (ELISAs) to monitor remediation and cleanup activities at Superfund sites will be developed. Each new method will be tested on real-world samples from monitoring studies. Field studies will be conducted when time and resources permit. The feasibility and application of immunosensors to provide field analytical methods for the dynamic monitoring of hazardous substances of interest to the EPA will also be investigated. Concern has been expressed for the potential exposure of individuals to toxic compounds who live near hazardous waste sites or who may become exposed through other means. Thus, the development of methods for measuring biomarkers for human exposure assessment studies is also addressed.
During the remainder of the Task several projects will be undertaken including:
- Complete the development and evaluation of bioanalytical methods for dioxin and related compounds
- Perform dioxin immunoassay analysis on samples from a dioxin SITE demonstration
- Comparison of an ELISA with gas chromatography (GC) for monitoring polychlorinated biphenyls (PCBs) in soils/sediments collected from a Superfund field demonstration
- Survey of bioanalytical methods and sensor technologies for environmental monitoring
- Development of immunoaffinity chromatography sample preparations for PCBs
- Preparation of standard operating procedures (SOPs) for each bioanalytical method developed
- Conduct yearly research meeting
- Conduct survey of high priority chemicals at National Priorities List (NPL) sites for bioanalysis suitability
- Develop new bioanalytical methods for hazardous compounds of public concern
- Perform PCP immunoassay analysis on soil and sediment samples from a Superfund site and compare with GC data
- Preparation of fact sheets and journal articles
Description:
The consumption of food commodities contaminated with the mold, Stachybotrys charatrum, has a long history of causing severe illness, and even death, in humans and animals. More recently, negative health effects have been linked to indoor inhalation exposures to S. chartarum. For these health effects to be documented, a mechanistic biomarker of exposure must be demonstrated. An enzyme linked- immunosorbent assay (ELISA) was developed for stachylysin, a proteinaceous hemolysin obtained from cultures of S. chartarum. The ELISA was highly selective to stachylysin and had an average limit of detection of 2 ppb depending upon the sample matrix. The conidia from several other molds showed a negligible reaction in the assay indicating the specificity of stachylysin to S. chartarum. The ELISA was used to analyze environmental samples from mold- contaminated buildings and biological samples obtained from individuals with known mold exposures. Stachylysin was quantified by the ELISA in spore, wallboard, mycelial and dust samples, and in the serum of adults exposed to S. chartarum in water-damaged environments. All control sera was negative for stachylysin. Thus, the analysis of stachylysin in serum may provide a useful measurement of human exposure to the mold. If stachylysin is a relevant biomarker, its determination in environmental samples may be useful for assessing and managing risk for indoor mold exposures.
Although this work was reviewed by EPA and approved for publication, it may not necessarily reflect official Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendation by EPA for use.