本文選題：城市醫(yī)院環(huán)境 + 空氣微生物　； 參考：《哈爾濱工業(yè)大學》2015年碩士論文

【摘要】：空氣中微生物與抗生素抗性基因(AntibioticResistanceGenes,ARGs)污染得到人們越來越多的關注。醫(yī)院因其場所的特殊性,存在著潛在的病原菌與ARGs污染問題。然而,目前對城市醫(yī)院環(huán)境空氣微生物與ARGs污染狀況的研究較少,缺乏人群在醫(yī)院環(huán)境暴露風險的基礎數(shù)據(jù)。本論文基于非培養(yǎng)的研究手段,建立了一套適合醫(yī)院空氣環(huán)境的微生物污染的檢測方法,并將該方法加以應用。論文研究了醫(yī)院環(huán)境空氣微生物群落結構、病原菌及ARGs污染狀況,基于該研究結果對人群在醫(yī)院環(huán)境的暴露狀況進行評估,以了解醫(yī)院空氣生物污染狀況,并初步探索了人群在醫(yī)院的暴露風險。研究建立了一種基于非培養(yǎng)的醫(yī)院空氣環(huán)境微生物污染的檢測方法。布點原則遵循空氣微生物監(jiān)測的標準,采樣方法為大流量TSP(TotalSuspendedParticulate)采樣器采集;采樣時間在22~26h之間;采樣膜預處理方法為酸性磁珠法;DNA樣品濃度的定量采用Qubit熒光法。對醫(yī)院空氣環(huán)境DNA樣品采用高通量測序技術和實時熒光定量PCR技術(qPCR)研究其空氣微生物的組成特征,病原菌及抗生素抗性基因的污染狀況�；�16SrRNA基因擴增片段的高通量測序結果表明:醫(yī)院空氣環(huán)境微生物的主導菌門為變形菌門、厚壁菌門等,空氣中病原菌的平均比例為3.64%,主導病原菌為腐生葡萄球菌、棒狀桿菌、大腸埃希氏菌等。典型醫(yī)院空氣的主導菌群和病原菌在冬夏季間差異明顯,病原菌比例夏季稍高于冬季;主導菌群的分布在空氣介質與塵土介質間略微不同,而病原菌比例塵土介質顯著高于空氣介質,表明塵土介質是醫(yī)院環(huán)境病原菌的一個潛在存儲庫。qPCR定量實驗的結果顯示:醫(yī)院空氣中β-內酰胺類ARGs相對濃度均值最大,與β-內酰胺類抗生素的使用率較高有關;其中β-內酰胺類的blaTEM基因與大環(huán)內酯類的ermB基因是醫(yī)院環(huán)境污染最嚴重的兩種基因;典型醫(yī)院四種ARGs(blaTEM、ermB、blaCTX-M、mecA)的污染在季節(jié)間差異不顯著,空氣介質中的相對濃度在103~107copies/ng范圍內,醫(yī)院環(huán)境空氣介質ARGs污染不容忽視。采用估算呼吸吸入潛在劑量值的方法來評估醫(yī)院環(huán)境空氣生物污染物暴露風險,結果顯示:病人每次輸液就診時可吸入細菌氣溶膠高于433CFU(Colony-FormingUnits),其中可入肺細菌氣溶膠高于145CFU,可入肺病原菌高于5CFU;門診部的醫(yī)護人員氣溶膠暴露風險最大;醫(yī)院環(huán)境ARGs暴露劑量為105~106copies/h,β-內酰胺類ARGs的暴露風險最大,且同種類ARGs暴露風險在不同部門間差別較小。
[Abstract]:More and more attention has been paid to the contamination of microorganism and antibiotic resistance gene antibiotic resilience genes in the air. Because of the particularity of the hospital, there are potential pathogens and ARGs pollution problems. However, there are few studies on microorganism and ARGs pollution in urban hospital environment, and lack of basic data on exposure risk of population in hospital environment. Based on the non-culture research methods, a set of methods for the detection of microbial contamination in hospital air environment were established and applied in this paper. The microbial community structure, pathogenic bacteria and ARGs pollution in hospital ambient air were studied in this paper. Based on the results of the study, the exposure status of the population in hospital environment was evaluated to understand the status of hospital air biological pollution. And preliminarily explored the exposure risk of the crowd in the hospital. A non-culture-based method for the detection of microbial contamination in hospital air environment was developed. The sampling method is a large flow rate TSP Total suspended Particulate sampler, the sampling time is between 22 ~ 26h, the sample film pretreatment method is the acid magnetic beads method and the Qubit fluorescence method is used to quantify the concentration of the sample. High-throughput sequencing technique and real-time fluorescence quantitative PCR technique were used to study the composition of airborne microbes and the contamination of pathogens and antibiotic resistant genes in hospital air environment DNA samples. The results of high throughput sequencing based on the amplified 16SrRNA gene showed that the dominant microbes in the air in hospital were Proteus, Thinacea, etc. The average proportion of pathogenic bacteria in the air was 3.64. The dominant pathogens were staphylococcus saprophyticus and Corynebacterium. Escherichia coli, etc. There were significant differences between the dominant bacteria and pathogenic bacteria in the air of typical hospitals during winter and summer, and the proportion of pathogenic bacteria in summer was slightly higher than that in winter, and the distribution of dominant bacteria was slightly different between air and dust. The proportion of pathogenic bacteria in dust medium was significantly higher than that in air medium, which indicated that dust medium was a potential repository of pathogenic bacteria in hospital environment. QPCR quantitative experiment showed that the average relative concentration of 尾-lactam ARGs in hospital air was the highest. Among them, 尾 -lactam blaTEM gene and macrolide ermB gene were two of the most serious environmental pollution genes in hospitals. The relative concentration of air media is in the range of 103~107copies/ng, and the ARGs pollution in hospital environment can not be ignored. The risk of exposure to biological pollutants in hospital ambient air was assessed by estimating the potential dose values of respiratory inhalation. The results showed that the inhalable bacterial aerosol was higher than 433 CFU colony in each transfusion visit, in which the pulmonary bacterial aerosol was higher than 145 CFU and the pulmonary pathogenic bacteria was higher than 5 CFU. The exposure dose of ARGs in hospital environment was 105 ~ 106copies-h. the exposure risk of 尾 -lactam ARGs was the highest, and the risk of exposure to the same type of ARGs was small among different departments.
【學位授予單位】：哈爾濱工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：X831

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