Development and application of quantitative bioaerosol analysis method using PCR

The presence of harmful airborne particulate matter of biological origin has been associated with variety of negative health effects. In addition, there is a real treat of malliciouse release of hazardouse bioaerosol to public sectors. To protect the population at risk from bioaerosol exposure, an effective bioaerosol detection system is urgently needed that enables a rapid and accurate bioaerosol sampling, identification, and quantification in air samples. As an effective bioaerosol monitorin system requires both an effective air sampling device and rapid sample analysis technique with high sensitivity, the performance of RCS High Flow was investigated with culture-based quantification technique in Chapter 1. The Results showed that the test sampler would collect more than 80 % of common fungal spores and more than 50 % of airborne bacteria larger than 1.1 mu m. However the biological performance of the sampler determined using a culturable bacterial counting method was significantly affected by environmental conditions, characteristics of sampler type, and consequently caused an underestimation in quantification. Therefore, in Chapter 2 and Chapter 3, Quantitative Real-Time Polymeratse Chain Reaction (QPCR) was applied to count the total bioaerol number in air samples. The results showed that successful bioaerosol quantification using QPCR requires not only to understand the characteristics of bioaerosol to be investigated and its sampling methodology, but also to develop study-specific standard curves. To increase the reliability of the method, the study-specific standrad curves associated with factors such as bacterial species, cell suspenssion preparation methods, QPCR methods should be developed and used for quantifications. To this end, the developed QPCR assay was applied to test the performance of a novel bioaerosol sampler (EPSS). The test results indicated a successful application of QPCR method to test performance of bioaerosol samplers. By coupling with an effective bioaerosol sampling device, this QPCR assay could increase the reliability of bioaerosol sampling systems and allow timely and effective quantification of aerosol samples. Overall, the findings in this dissertation provide the general guidelines to develop an effective bioaerosol monitoring system by setting-up the study-specific protocol of QPCR assay capable of determining total cell numbers in air samples. The improved bioaerosol sampling system enabling rapid quantification of bioaerosols with high sensitivity may be applied as a basis for developing bioaerosol detection systems capable of detecting even small bioaerosol concentrations thus providing useful information needed to understand the bioaerosol exposure dose and response relationship.