即時檢測沙門氏桿菌及其抗藥性
之適體傳感器

在亞洲地區當中，沙門氏菌是幼兒腸道炎常見的病菌，而現行檢測沙門氏菌抗藥性的塗盤方式需耗費三至五天的時間，若能縮短檢測抗藥性的時間，不但能夠幫助醫師選擇適當的抗生素，也可以避免抗生素濫用的情形。本研究的目的是發展一種可即時檢測菌體存在，並在數小時內完成細菌抗生素篩選的適體傳感器。採用新式奈米材料－單壁奈米碳管，使其鍵結與沙門氏桿菌具有專一性之核酸適體，並利用微機電製程技術，製作以鉻金屬為材料之平行梳狀電極，再使用微量吸管吸取沙門氏桿菌液滴定於載具容器的電極上，讓其與核酸適體接合產生電荷傳遞，並可直接於電極上做細菌的培養。當電極上的細菌隨著時間的增生，電訊號也會因此改變，最後加入抗生素於培養載具中，使用微電流計與阻抗分析儀擷取電訊號，紀錄其訊號變化差異性。本研究得到馬偕醫院檢驗科的協助，提供實驗用的標準菌，包括大腸桿菌、沙門氏桿菌、檸檬酸桿菌、以及變形桿菌等不同的腸內菌種來測試晶片的專一性，並藉由沙門氏菌增生而改變的電流變化率，來判斷菌種是否具有抗藥性，這種簡易的判讀方式有助於臨床實驗的操作，未來修飾更多細菌種類的核酸適體晶片，即可為一種細菌檢驗的使用平台。

關鍵字

微機電製程技術；單壁奈米碳管；核酸適體；抗藥性檢測

並列摘要

The development of an aptamer-based sensor for typing of bacteria is presented. Highly specific DNA aptamers to Salmonella enteritidis were selected via Cell-SELEX technique. This study developed a method of detecting bioparticles such as Salmonella by a biochip using single-walled carbon nanotubes modified by aptamer. A label-free impedance sensor for rapid detection of Salmonella was developed by aptamer onto an chromium interdigitated array (IDA) microelectrode. The cross-test between Salmonella and E. coli on the biochip exhibits high specificity for Salmonella. Each comprises a positive selection step against Salmonella enteritidis and a negative selection step against a mixture of related pathogens, including Salmonella, Escherichia coli, Citrobacter freundii, and Proteus to ensure the species-specificity of the selected aptamers. The electron-transfer resistance was correlated with the concentration of Salmonella. A variety of different antibiotics were dropped in the wells with volume of 0.5 ml on the biochip to detect antibiotics resistance of Salmonella.

並列關鍵字

MENS ； Carbon Nanotube ； Aptamer ； The drug resistance of bacteria

參考文獻

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[7] H.A. Crissman and J.A. Steinlamp, ”Rapid simultaneous measurement of DNA, protein and cell volume in single cells from large mammalian cell populations,” J.Cell Biol, Vol 59, 1973, pp.766.

[9] J. R. Webster, M. A. Burns, D. T. Burke, C. H. Mastrangelo, “Monolithic Capillary Electrophoresis Device with Integrated Fluorescence Detector,” Anal. Chem, 2001, pp.1622-1626.

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即時檢測沙門氏桿菌及其抗藥性之適體傳感器

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