Evaluation of PCR interference from dust samples for monitoring indoor environments.
Keswani J; Chen B
Abstracts of the 2002 Annual Conference of the American Society for Microbiology, May 19-23, 2002, Salt Lake City, Utah. Washington, DC: American Society for Microbiology, 2002 May; :452-453
Viable and non-viable biological contaminants in the indoor workplace have been reported to cause health problems ranging from allergic and infectious diseases to nonspecific "sick building symptoms". Recently, Polymerase Chain Reaction (PCR) technology has gained importance as a diagnostic tool for rapid detection and characterization of microorganisms. However, the potential interference with the PCR due to the presence of organic and inorganic compounds in die environmental samples have lately drawn much attention in this area. Although progress has been made by including DNA purifications in the conventional PCR protocol to reduce the interference, no detailed study has been reported in the literature by using a simple PCR protocol on fungal spores. The focus of this research was to investigate the practicality of applying a simple PCR method to the field with minimal manipulations to maintain the integrity of samples. A total of four real-life samples collected from a poultry farm, two hospital rooms and HVAC filters were studied in detail to investigate interference on PCR. The dust samples were spiked with known numbers of Stachybotrys chartarum spores, ranging from 2 x 102 to 2x 105. The method of bead-beating was used for disruption of spores and release of DNA.
Dust particles; Dust sampling; Dusts; Monitors; Health hazards; Infectious diseases; Allergies; Microorganisms; Organic compounds; Inorganic compounds; Environmental contamination
Research Tools and Approaches: Exposure Assessment Methods
Abstracts of the 2002 Annual Conference of the American Society for Microbiology, May 19-23, 2002, Salt Lake City, Utah