利用奈米金粒子結合拓印法來偵測培養皿上的沙門氏菌

Colony-printing gold-nanoparticle immunoassay in detection of Salmonella spp. isolated on media

曾瑋佑(Wei-Yu Tzeng)

指導教授 ： 侯劭毅

國立臺北科技大學/工程學院/生物科技研究所/碩士(2008年)

https://doi.org/10.6841/NTUT.2008.00508

摘要

本研究主要是發展一套非PCR類的、並且能夠快速偵測細菌的方法；這樣較快速的方法能夠及早發現病原菌的存在並且幫助臨床醫生做出適當的決策。所以在本實驗中，我們利用接有抗沙門氏菌抗體奈米金粒子以及拓印法兩者結合以應用於偵測醫院臨床上的樣本中，是否含有沙門氏菌存在。拓印法利用硝化纖維膜在長有沙門氏菌及其它的菌種的瓊脂平板上，在一定範圍內輕輕沾粘菌落，如此一來，膜上除了黏有菌外，各種菌落的相對位置也被記錄了下來，之後再將此膜浸泡於接有沙門氏菌抗體、紅色奈米金粒子溶液中，結果欲偵測的菌種就會在膜上顯示出顏色，其它菌種則不會呈現顏色，之後觀察膜上有顏色的部份，利用其相對位置，比照一下瓊脂平板上生長的菌落，即可知哪一個菌落是屬於沙門氏菌的菌。本研究從2007年9月到2008年4月從台北馬偕醫院收集病人檢體，總數為577個，將樣本培養於HE plate上並做檢測；測試結果與Vitek 2測試結果做比較，發現本研究檢測技術靈敏度可高達100%，專一性高達99%左右。

關鍵字

沙門氏菌 ； 奈米金粒子 ； 抗體 ； 拓印法

並列摘要

This study is to develop a non-PCR method for early detection of bacteria which can lead to help clinical physicians in making proper decisions. Different bacteria were spread on agar plates and then incubated to form colonies. A dry nitrocellulose paper was used to gently cover the agar surface by a one-touch press only by the paper’s own weight. The protocol was based on the transfer of surface cells of the colonies to the nitrocellulose membrane. Gold nanoparticles coated with the anti-Salmonella antibody were used to highlight the Salmonella on the membrane to facilitate the selectivity. The red dots on the membrane indicate the existence of Salmonella because gold nanoparticles are red instead of golden. From September 2007 to April 2008, 577 stool samples from patients at Mackay Memorial Hospital, Taipei were tested on Hektoen enteric agar plate. The black or crystalloid colonies were tested using this method. Compared with Vitek 2, the sensitivity of this new method is 100% and the specificity is about 99%.

並列關鍵字

Salmonella ； gold-nanoparticle ； antibody ； Colony-printing

參考文獻

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