Monitoring, quantification and invasion assessment of Legionella in hospital water sources by culture, real-time PCR and cell culture

سال انتشار: 1399
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 181

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شناسه ملی سند علمی:

MEDISM21_013

تاریخ نمایه سازی: 23 مرداد 1400

چکیده مقاله:

Background and Aim : Legionella is a heterophilic bacterium in the water supply network, which can normally survive in the biofilm or planktonic form in freshwater and is an optical intracellular bacterium and will be found only after proliferating within macrophages and rip them out in large numbers in the alveolar spaces and the lower respiratory secretions. the bacteria enter the lungs through the formation of aqua aerosol and contamination with Legionella species includes the spectral of disease that is the Pontiac fever, self-limited influenza-like disease in one hand, and systemic and legionnaire's disease depends on the contamination of water sources and susceptibility. The aim of this study was to Using culture, quantitative PCR for Legionella monitoring in hospital water samples, and risk assessment by an invasion of the HeLa cell.Methods : In this study ۱۰۰ water samples collected from hospitals of Iran university medical sciences, from head-showers, humidifier bottles, and laboratory water baths and were examined. To prevent the growth of fast-growing bacteria and saprophytic fungi, all samples were treated with acid. Then they were cultures on selective media such as GVPC. To confirm the primary identity of the isolates colonies, gram staining, and lack of growth after insemination to conventional laboratory environments such as blood agar and BCYE without L-CYS. DNA of the samples was extracted, and then the PCR reaction for ۱۶srRNA genes was done using proprietary primers. Eventually, after electrophoresis an determination of PCR products, they were analyzed. After that real-time PCR to measuring the number of bacterial was done and invasion of isolates was asses by the HeLa cell culture.Results : Out of ۱۰۰ water samples were collected, ۱۲ cases (۱۲%) were positive by culture method and ۴۲ cases (۴۲%) were positive by the PCR method, and results of culture were confirmed. Quantification of bacteria in ۲۳ cases (۵۴/۵۶%) was with ۱۰۴GU/L and ۲ cases (۴/۷۶%) were with <۱۰۴ GU/L. Finally, the invasion of isolates was assessed by the HeLa cell culture.Conclusion : Despite the use of refined water resources for the urban distribution system, ۱۲% of the samples taken from hospital water sources were contaminated with legionella bacterium; on the other hand, because the legionella bacterium was resistant to disinfectant such as chlorine with common concentration.

کلیدواژه ها:

Legionella - hospital water sources - hela cell culture - real time PCR

نویسندگان

Shirin Bavari

Department of microbiology - Faculty of medicine - Iran university of medical sciences

Shiva Mirkalantari

Department of microbiology - Faculty of medicine - Iran university of medical sciences