Detection of Pathogenic Bacteria in the Air by Culture Techniques in Combination with Multiplex Polymerase Chain Reaction
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Abstract
Respiratory infections are among important infections in human. The causative agents are diverse group of bacteria, virus, fungi and parasites. This research focuses on the detection of microbial contaminants and respiratory pathogens in indoor air of university environment by application of a molecular method. Seven pathogens including Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, Pseudomonas aeruginosa and Legionella pneumophila were screened. Twenty-eight air samples were analyzed by conventional culture method and the multiplex polymerase chain reaction (mPCR) that detected those seven pathogens simultaneously within three reactions. The conventional culture method showed the average total count of large lecture rooms at 3.4 CFU plate-1 h-1, small lecture rooms at 5.2 CFU plate-1 h-1, meeting rooms at 4.3 CFU plate-1 h-1, office rooms at 3.0 CFU plate-1 h-1, canteen rooms at 31.5 CFU plate-1 h-1 and service vans at 5.5 CFU plate-1 h-1. Contaminants comprised Gram-positive bacteria, Gram-negative bacteria and molds. The genus compositions were Staphylococcus spp. (62.50%), Micrococcus spp. and Kytococcus spp. (58.33%), Bacillus spp. (41.64%), Moraxella spp. (29.17%), Corynebacterium spp. (25.00%) and Pseudomonas spp. (20.83%). MultiplexPCR was able to detect Staphylococcus aureus and Streptococcus pyogenes at 16.67% and 4.17% of air samples, respectively, whereas conventional method did not. These two bacteria are important pathogens of human and are common cause of wide range of infections. The results suggested that mPCR was a useful supplement to the conventional method to monitor indoor air microbiological quality and should be introduced in guidelines of indoor air quality monitoring.
Keywords: Airborne bacteria; respiratory infections; multiplex polymerase chain reaction
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E-mail: chanchaemw@yahoo.com
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