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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/26974

Title: Rapid Diagnostics of Aquaculture Fish Pathogen Edwardsiella tarda by the Use of a Simple Designed DNA Hybridization Microfluidic Device
Authors: 陳昭德;陳柏台;張忠誠
Jau-Der Chen;Pei-Tai Chen;Chung-Cheng Chang
Contributors: NTOU:Department of Aquaculture
Keywords: ET996;Edwardsiella tarda;Microfluidic chip;Soft lithography;16S rDNA specific probe ET996
Date: 2010-03-01
Issue Date: 2011-10-21T02:20:33Z
Publisher: Journal of the Fisheries Society of Taiwan
Abstract: Abstract:A micro-channel biochip is made for rapid discrimination of probe-DNA hybridization. The chip is designed by comprising a hybridization room inside with a porous nylon membrane and connecting with two flow-through aperture as inlet and outlet. Bacterial DNA is directly injected into the positively charged nylon membrane from the inlet of the chip. The DNA is fixed and denatured on the membrane at 120℃ for 20 minutes. The 20-mer oligonucleotides ET996-HEX which specify to 16S rDNA of Edwardsiella tarda are added for probe-DNA hybridizing at 68℃ for 5 minutes and then cooled spontaneously at room temperature. Then, a low salt buffer at a constant speed 15 μl/min is pumped into the polydimethylsiloxane chip to wash the membrane and elute the un-hybridized fluorescent probe out of the chip at room temperature. The residues of the fluorescent probe on the membrane of the chip are detected simultaneously by a photo-sensor. The success of DNA-probe hybridization can be determined within 10 minutes after buffer elution. Our work establishes a rapid E. tarda species identifying platform for earlydiagnostics and is potential in preventing from epidemic outbreaks.
摘要:製作一微流道生物晶片用來快速區別核酸與探針之雜合,該晶片設計具有一雜合室,室內置有一帶有孔洞之尼龍膜,及兩條微流道做為與外連結之進出口。細菌之核酸直接由晶片流道進口注入,經由微流道到達帶有正電荷的尼龍膜上,以120℃高溫烤20分鐘,改變核酸的性質固定於尼龍膜上,再注入愛德華氏菌小次單位核醣體帶有螢光的專一性探針ET996,於68℃雜合5分鐘並在室溫下自然降溫,然後在室溫下以低鹽洗液定速(15 μl/min)沖洗尼龍膜,將未雜合之帶有螢光的探針ET996,經由另一微流道出口洗出晶片外,殘餘在微流道晶片上的螢光探針,同步以光感測器偵測,核酸與探針的成功雜合,可於洗液沖洗10分鐘內決定。
Relation: 37(1), pp.65-76
URI: http://ntour.ntou.edu.tw/handle/987654321/26974
Appears in Collections:[水產養殖學系] 期刊論文
[電機工程學系] 期刊論文

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