環保專案成果報告查詢系統

專案計畫編號	EPA-100-U1U4-04-001
經費年度	100
計畫經費	1,638 千元
專案開始日期	2011/03/22
專案結束日期	2011/11/30
專案主持人	陳中庸
主辦單位	永續發展室
承辦人	林燕柔
執行單位	中原大學
專案分類	環境監測及檢測
中文關鍵字	退伍軍人菌、量子點、專一性單股核酸探針
英文關鍵字	Legionella pp.、Quantum dots、Single oligonucleotide specific probes
協同主持人	吳宗遠
共同主持人
計畫聯絡人	陳中庸
計畫聯絡電話	03-2653540#
計畫聯絡信箱	cychen@cycu.edu.tw

中文摘要	本「開發奈米材料量子點應用於環境致病性微生物之生物晶片」計畫為開發一新的微生物檢測技術並同時解決定量與標定問題，且期望簡化檢體製備流程。利用量子粒的特性配合微珠嵌連之專一性單股核酸探針，提供一全新技術解決上述問題的方向。環境致病性微生物中退伍軍人菌一直是公衛的重點，因為像醫院、旅館、室內商場、辦公大樓、三溫暖等大型建築物的水塔均是可能的汙染源。然而退伍軍人症的臨床檢驗採細菌培養，費時且易錯失治療的黃金時機，因此發展一套精確、快速的基因檢測方法有其重要性與必要性。我們目前以針對8種常見之退伍軍人菌株完成 1) 8種專一性單股核酸序列之設計與製備。 2) 16種專一性探針之設計與製備。 3) 將8種專一性單股核酸序列接合到微珠。 4) 將16種專一性探針接合到量子粒。 5)完成8種退伍軍人菌株之DNA 檢體製備。6)完成探針穩定測試，於室溫下穩定超過1個月。7) 此新偵測法達到 1ng 的偵測極限。8) 此新偵測法達到R2=0.984 的標準偵測曲線。9) 此新偵測法敏感度高無偽陽性。10) 此新偵測法專一性達100%，無偽陰性。本研究提供一新有效偵測法，可同時節省偵測時間又可定量。
中文關鍵字	退伍軍人菌、量子點、專一性單股核酸探針

中文摘要

本「開發奈米材料量子點應用於環境致病性微生物之生物晶片」計畫為開發一新的微生物檢測技術並同時解決定量與標定問題，且期望簡化檢體製備流程。利用量子粒的特性配合微珠嵌連之專一性單股核酸探針，提供一全新技術解決上述問題的方向。環境致病性微生物中退伍軍人菌一直是公衛的重點，因為像醫院、旅館、室內商場、辦公大樓、三溫暖等大型建築物的水塔均是可能的汙染源。然而退伍軍人症的臨床檢驗採細菌培養，費時且易錯失治療的黃金時機，因此發展一套精確、快速的基因檢測方法有其重要性與必要性。我們目前以針對8種常見之退伍軍人菌株完成 1) 8種專一性單股核酸序列之設計與製備。 2) 16種專一性探針之設計與製備。 3) 將8種專一性單股核酸序列接合到微珠。 4) 將16種專一性探針接合到量子粒。 5)完成8種退伍軍人菌株之DNA 檢體製備。6)完成探針穩定測試，於室溫下穩定超過1個月。7) 此新偵測法達到 1ng 的偵測極限。8) 此新偵測法達到R2=0.984 的標準偵測曲線。9) 此新偵測法敏感度高無偽陽性。10) 此新偵測法專一性達100%，無偽陰性。本研究提供一新有效偵測法，可同時節省偵測時間又可定量。

中文關鍵字

退伍軍人菌、量子點、專一性單股核酸探針

類型	檔名	檔案大小	說明	最新版公開日期
期末報告	開發奈米材料量子點應用於環境致病性微生物之生物晶片-期末報告(公開版).pdf	0MB		2012/3/1 上午 12:00:00

摘要	The main purpose of this project is to adopt quantum dots and to generate new technology for detection of bacterial pathogen. The new technology aims to measure the quantity and quality by simple labeled methods thus the detection procedure will be short and faster than current methods. We propose the quantum dot probes combined with microbead linked specific oligonucleotides as detection system that will be able to provide higher sensitivity and specificity. We selected Legionella spp as our research targets. Legionella spp has been pointed out as the major environmental pollution issue by CDC of government. Legionnaires’ disease (LD) is caused mainly by inhalation of aerosols generated from water sources contaminated with Legionella spp. Environmental culturing of water systems for Legionella spp. has been recommended for hospitals. Thus, we developed our quantum dots based microbeads system for 8 common Legionella spp. Currently, we have already designed and obtained oligonucleotides mcirobead and quantum dot probes for 8 Legionella spp. Those 8 individual oligonucleotides have been linked into microbeads and those 16 specific probes have been conjugated to quantum dots. Those quantum dots probes can stand up to one month at room temperature. Our novel detection system can detect lower as 1ng of pathogen DNA. Moreover, the standard curve of this system can reach R2=0.984 with high sensitivity and 100% of specificity. Therefore, we have developed a useful tool to detect pathogen with shorter operation time and to quantify the pathogen DNA in high specificity and sensitivity.
英文關鍵字	Legionella pp.、Quantum dots、Single oligonucleotide specific probes

摘要

The main purpose of this project is to adopt quantum dots and to generate new technology for detection of bacterial pathogen. The new technology aims to measure the quantity and quality by simple labeled methods thus the detection procedure will be short and faster than current methods. We propose the quantum dot probes combined with microbead linked specific oligonucleotides as detection system that will be able to provide higher sensitivity and specificity. We selected Legionella spp as our research targets. Legionella spp has been pointed out as the major environmental pollution issue by CDC of government. Legionnaires’ disease (LD) is caused mainly by inhalation of aerosols generated from water sources contaminated with Legionella spp. Environmental culturing of water systems for Legionella spp. has been recommended for hospitals. Thus, we developed our quantum dots based microbeads system for 8 common Legionella spp. Currently, we have already designed and obtained oligonucleotides mcirobead and quantum dot probes for 8 Legionella spp. Those 8 individual oligonucleotides have been linked into microbeads and those 16 specific probes have been conjugated to quantum dots. Those quantum dots probes can stand up to one month at room temperature. Our novel detection system can detect lower as 1ng of pathogen DNA. Moreover, the standard curve of this system can reach R2=0.984 with high sensitivity and 100% of specificity. Therefore, we have developed a useful tool to detect pathogen with shorter operation time and to quantify the pathogen DNA in high specificity and sensitivity.

英文關鍵字

Legionella pp.、Quantum dots、Single oligonucleotide specific probes