Abstract
摘要:退伍軍人菌與棘阿米巴原蟲同屬環境傳播病原菌,感染後皆可導致人類死亡。從公共衛生觀點,若能實施適當環境監測技術,將有助於掌握民眾暴露風險,進而探究影響暴露風險之因子,以俾採取對策以維護大眾健康。目前已有可準確快速定量環境中活性與總退伍軍人菌以及總棘阿米巴原蟲之分析方法;但在定量活性棘阿米巴原蟲方面,仍尚有諸多分子生物技術議題(如:核酸染劑種類、染劑濃度、光活化條件、偵測限值等)有待研究予以釐清。此外,退伍軍人菌導致的致死性肺炎(亦即退伍軍人病)多屬零星感染,且以職業駕駛之得病風險較高。對此,雖然近期流行病學研究指出其可能與未使用車輛雨刷清潔劑有關,然目前尚無詳盡的環測調查得予佐證,也欠缺足夠的科學證據可釐清導致車體遭退伍軍人菌污染之主要貢獻因子。 有鑑於此,本計畫將以三年為期探究上述議題。第一年將透過實驗室開發與環境驗證方式,建立可快速且正確定量活性棘阿米巴原蟲之分子生物學方法。第二年將進行多類型車輛與退伍軍人菌污染之研究,針對職業與非職業駕駛所用車輛之空調與雨刷水系統進行採樣分析,並紀錄用車行為及車體清潔維護狀況;此外,也將測量可能影響退伍軍人菌滋生於雨刷水之多項水質因子。由於活性棘阿米巴原蟲是退伍軍人菌之自然宿主,可促使退伍軍人菌的增生,故於第三年將以本計畫建立之活性棘阿米巴原蟲定量技術,分析車體樣本中活性原蟲濃度,並將此數據與其他收集之車體資訊、用車行為、雨刷水與空調系統操作與維護方式、以及各類水質因子數據進行單變項與多變項統計分析,以解析顯著影響總、活性以及可培養退伍軍人菌滋生之因子。 預期本計畫所獲致之結果,將可釐清車體退伍軍人菌之污染來源、污染程度及其顯著影響因子,提供學術界現所欠缺之科學證據與知識。而此知識也將有助於研擬控制車體污染之管理對策。據此,透過降低車輛駕駛、搭車民眾以及周遭用路人暴露於病原菌之風險,達到落實公共衛生預防疾病之目的。此外,本研究所建立之活性棘阿米巴原蟲定量技術,未來也可應用於其他設施與環境,以評估棘阿米巴原蟲可引發的疾病風險及其對其他寄生病原細菌(如:退伍軍人菌、綠膿桿菌、分枝桿菌)族群流佈之影響。
Abstract: Legionella and Acanthamoeba are environmental pathogens, which may cause death in humans. From the point of view of public health, quantifying the population of these two pathogens in environments by adopting appropriate monitoring techniques is essential in order to well understand the risk posed to human health and to explore the factors affecting their distribution. Suitable monitoring techniques have been developed for Legionella; however, the optimal method for quantifying viable Acanthamoeba remains unsolved. In addition, lethal pneumonia caused by Legionella (i.e. Legionnaires' disease) occurs mainly as sporadic cases and is associated with occupational drivers. Although epidemiological data suggest a link to non-use of screen wash in vehicle, thorough environmental surveillance and the research to explore the factors significantly contributing Legionella contamination in vehicle are unavailable. To address these issues, a three-year project will be conducted by first developing the analytical technique for accurate quantification of viable Acanthamoeba via laboratory and environmental evaluation. The second year will begin with the assessment of Legionella contamination in vehicle. Various water quality parameters will be measured from windshield wiper fluids and the information about the vehicle use and maintenance will also be obtained via questionnaire. Considering Acanthamoeba are the natural hosts of Legionella, the technique developed for viable Acanthamoeba in this project will be adopted to quantify Acanthamoeba in vehicle samples. Acanthamoeba data will be included with other collected information to determine the statistically significant factors influencing Legionella contamination in vehicles in the third year. It is expected that this project will provide scientific evidence and new knowledge about the source and level of Legionella contamination in vehicle and their significant contributing factors. This knowledge will be helpful in planning control measures to minimizing microbial contamination, so that the drivers, passengers and the people on the road may be prevented from acquiring infectious diseases caused by Legionella and Acanthamoeba. Moreover, the analytical assay developed for viable Acanthamoeba may be applied to various environments, and the information obtained will significantly improve our knowledge about the exposure risk of Acanthamoeba and their impact on their bacterial parasites (e.g., Legionella, Pseudomonas and Mycobacterium).
Keyword(s)
退伍軍人菌
棘阿米巴原蟲
車輛
ethidium monoazide
propidium monoazide
定量聚合酶連鎖反應
Legionella
Acanthamoeba
vehicles
ethidium monoazide
propidium
monoazide
quantitative polymerase chain reaction