Study On Fast And Quantitative Detection Method Of Three Common Pathogenic Airborne Bacteria

The airborne microorganisms are not only the source of contamination in the microbial lab,but also make up of potential threats to the health of the work staff. Culture-independentmethods were used to analysis the composition and diversity of the airbrone bacteria, thenquantitative detection procedures for Escherichia coli, Staphylococcus aureus andAcinetobacter baumanii were established respectively on the basis of16S rDNA clone libraryanalysis.In this study cellulose membranes were used to collect the indoor airborne bacteria. Differentschemes were devised according to the pore size of the cellulose membrane, speed of airflowand the time collected. The result revealed that the optimal pore size was0.22μm. Because ofthe tiny effect of the time collected and the speed of airflow on the collection of airbornebacteria, enough collection time should be guaranteed according to different speed of airflowwhen collecting airborne bacteria. In this study a comparison was made among differentmethods of DNA extraction. The result revealed that the effect of the modified Soil genomicDNA fast isolation kit (BioTeke Corporation) is the best. The composition and diversity of theairborne bacteria was analyzed by the cloned16S rDNA library. The cloned library containedBacillus (60.3%), Acinetobacter (19.4%), Paracoccus (14.9%), Staphylococcus (1.2%),Enterobacter (0.7%), Corynebacterium (0.7%), Cupriavidus (0.5%), Schlegelella (0.5%),Sphingopyxis (0.5%), Xanthomonadaceae (0.5%), Microvirga (0.2%), Sphingomonas (0.2%),uncultured Pseudomonadales (0.2%) and uncultured Rhodobacterales (0.2%). Then threecommon pathogenic bacteria, Escherichia coli, Staphylococcus aureus and Acinetobacterbaumanii, were chosen for quantitative detection study. The specific primers with highcoverages of Escherichia coli, Staphylococcus aureus and Acinetobacter baumanii weredesigned and screened using BioEdit, primer premier5.0, DNAStar, DNAUser andAnnbyb222, and checked by PCR and real time quantitative PCR procedure. And the standardprocedures and standard curves for quantitative fluorescent PCR detection of these threecommon airborne pathogenic bacteria were established.