環保專案成果報告查詢系統

專案計畫編號	EPA-98-E3S5-02-01
經費年度	098
計畫經費	1,480 千元
專案開始日期	2009/02/05
專案結束日期	2009/12/31
專案主持人	張長泉
主辦單位	環檢所
承辦人	許令宜
執行單位	國立成功大學醫學檢驗技術研究中心
專案分類	環境監測及檢測
中文關鍵字	核糖體核酸基因內轉錄區、寡核苷酸晶片、空氣致病菌
英文關鍵字	Internal transcribed spacer, oligonucleotide array, air-borne fungi
協同主持人
共同主持人
計畫聯絡人	蘇淑琍
計畫聯絡電話	06-2353535#5790
計畫聯絡信箱	shuli0827@yahoo.com.tw

中文摘要	台灣位處亞熱帶，氣候溫暖潮濕極適合真菌的生長。飄浮在空氣中的黴菌孢子或代謝物，易成為人類的過敏原，引發支氣管哮喘、蕁麻疹、過敏性鼻炎、結膜炎、腸胃炎、皮膚炎等過敏症，如能及早偵測到這些真菌存在空氣中，應能採取管制，降低風險。傳統真菌鑑定方法往往需要專業的微生物人才與漫長分析時間，開發一套環境中致病真菌的生物檢測晶片，可以同時監測多種致病真菌，兼具簡單、快速與正確的特色。為快速監測空氣中有害黴菌的存在，本研究發展寡核苷酸晶片以鑑定空氣中有害黴菌，利用核糖體核酸基因內轉錄區(internal transcribed spacer, ITS)設計特異性之寡核苷酸探針，並將其點在尼龍膜上製成晶片，待測菌種經PCR放大ITS區域後與晶片上之探針進行雜合反應。本晶片可鑑定21種有害黴菌，總共測試139株菌（73株目標菌株及66株非目標菌株），鑑定率為98.6% (72/73)，特異性(specificity)為100% (66/66)。進一步利用此晶片實測空氣樣本的分離株143株，某政府單位樣本由晶片可檢測到Alternaria alternata, Aspergillus flavus, Aureobasidium pullulans, Cladosporium cladosporides。某醫院及某研究大樓樣本由晶片可檢測到Aspergillus niger, Aspergillus versicolor，晶片鑑定率為100% (71/71)，特異性為98.6% (71/72)。由目前結果顯示此晶片為一種可鑑定空氣中有害黴菌之良好工具。
中文關鍵字	核糖體核酸基因內轉錄區、寡核苷酸晶片、空氣致病菌

中文摘要

台灣位處亞熱帶，氣候溫暖潮濕極適合真菌的生長。飄浮在空氣中的黴菌孢子或代謝物，易成為人類的過敏原，引發支氣管哮喘、蕁麻疹、過敏性鼻炎、結膜炎、腸胃炎、皮膚炎等過敏症，如能及早偵測到這些真菌存在空氣中，應能採取管制，降低風險。傳統真菌鑑定方法往往需要專業的微生物人才與漫長分析時間，開發一套環境中致病真菌的生物檢測晶片，可以同時監測多種致病真菌，兼具簡單、快速與正確的特色。為快速監測空氣中有害黴菌的存在，本研究發展寡核苷酸晶片以鑑定空氣中有害黴菌，利用核糖體核酸基因內轉錄區(internal transcribed spacer, ITS)設計特異性之寡核苷酸探針，並將其點在尼龍膜上製成晶片，待測菌種經PCR放大ITS區域後與晶片上之探針進行雜合反應。本晶片可鑑定21種有害黴菌，總共測試139株菌（73株目標菌株及66株非目標菌株），鑑定率為98.6% (72/73)，特異性(specificity)為100% (66/66)。進一步利用此晶片實測空氣樣本的分離株143株，某政府單位樣本由晶片可檢測到Alternaria alternata, Aspergillus flavus, Aureobasidium pullulans, Cladosporium cladosporides。某醫院及某研究大樓樣本由晶片可檢測到Aspergillus niger, Aspergillus versicolor，晶片鑑定率為100% (71/71)，特異性為98.6% (71/72)。由目前結果顯示此晶片為一種可鑑定空氣中有害黴菌之良好工具。

中文關鍵字

核糖體核酸基因內轉錄區、寡核苷酸晶片、空氣致病菌

類型	檔名	檔案大小	說明	最新版公開日期
期末報告	計畫期末報告全文-EPA-98-E3S5-02-01.pdf	3MB		2010/5/1 上午 12:00:00

摘要	Fungi exist in both indoors and outdoors. Exposure to fungi, particularly in water damaged indoor environments, has been thought to exacerbate a number of adverse health effects, ranging from subjective symptoms such as fatigue, cognitive difficulties or memory loss to more definable diseases such as allergy, asthma and hypersensitivity pneumonitis. Traditionally, fungi have been recovered by plate cultivation. Fungal identification may require days to weeks and expertise is needed for reliably identification. Recently, DNA microarray technology displays a high potential in the testing and analysis of environmental samples. Recent applications of the DNA microarray in environment studies include the detection of pathogens, environmentally functional genes, specific bacteria and viruses. In this study, based on the sequences of the ITS regions, an oligonucleotide array was developed to identify fungal pathogens in air. A collection of 73 target strains (21 species) were analyzed by the array and a sensitivity of 98.6% (72/73) was obtained. In addition, a total of 66 nontargrt strains (66 species) were also tested and a specificity of 100% (66/66) was obtained. Furthermore, 143 isolates from air samples were analyzed by the array; the correct identification rate was 100% (71/71) and the specificity was 98.6% (71/72). The present method is a rapid and accurate alternative for identification of fungi isolated from air samples.
英文關鍵字	Internal transcribed spacer, oligonucleotide array, air-borne fungi

摘要

Fungi exist in both indoors and outdoors. Exposure to fungi, particularly in water damaged indoor environments, has been thought to exacerbate a number of adverse health effects, ranging from subjective symptoms such as fatigue, cognitive difficulties or memory loss to more definable diseases such as allergy, asthma and hypersensitivity pneumonitis. Traditionally, fungi have been recovered by plate cultivation. Fungal identification may require days to weeks and expertise is needed for reliably identification. Recently, DNA microarray technology displays a high potential in the testing and analysis of environmental samples. Recent applications of the DNA microarray in environment studies include the detection of pathogens, environmentally functional genes, specific bacteria and viruses. In this study, based on the sequences of the ITS regions, an oligonucleotide array was developed to identify fungal pathogens in air. A collection of 73 target strains (21 species) were analyzed by the array and a sensitivity of 98.6% (72/73) was obtained. In addition, a total of 66 nontargrt strains (66 species) were also tested and a specificity of 100% (66/66) was obtained. Furthermore, 143 isolates from air samples were analyzed by the array; the correct identification rate was 100% (71/71) and the specificity was 98.6% (71/72). The present method is a rapid and accurate alternative for identification of fungi isolated from air samples.

英文關鍵字

Internal transcribed spacer, oligonucleotide array, air-borne fungi