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首頁> 外文學(xué)位 >Development of capillary electrophoresis-based microfluidic devices for future medical diagnostics: Separation and detection of DNA adducts and other related biomarkers.
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Development of capillary electrophoresis-based microfluidic devices for future medical diagnostics: Separation and detection of DNA adducts and other related biomarkers.

機(jī)譯：基于毛細(xì)管電泳的微流控設(shè)備的開發(fā)，可用于未來的醫(yī)學(xué)診斷：分離和檢測(cè)DNA加合物和其他相關(guān)生物標(biāo)記。

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Genotoxic carcinogens can react covalently with DNA to form DNA adducts, which can lead to mutations in critical genes and subsequently to the development of diseases such as cancer. Consequently, the role of DNA damage and their subsequent biomarkers are considered important in studies involving cancer and other aging-related diseases. Analytical methods with high resolving power and sensitive detection are needed to detect neurotransmitters (e.g. dopamine (DA) and DA-derived DNA adduct), catechol estrogen-derived DNA adducts, and 8-hydroxy-deoxyguanosine DNA adduct in human fluids, as their presence is difficult to determine by standard chromatography with UV absorbance detection. Furthermore, these DNA adducts are weakly fluorescent at room temperature. Thus, native fluorescence detection methods are ineffective with this class of biomarkers. Interestingly, capillary electrophoresis (CE) combined with electrochemical detection (ED) has been found to be a sensitive, selective technique for analyzing a large number of compounds including these analytes. Therefore, a CE-based PDMS/glass hybrid microfluidic device for free solution electrophoresis with totally integrated gold electrodes and simplified gated sample injection were fabricated to separate and detect various DNA adducts and other related biomarkers. Various arrangements of the ED system were tested and evaluated including in-channel and end-channel detection. The best electrode configuration was optimized based on in-channel detection with an in situ fabricated palladium decoupler serving as the electrophoretic ground located inside the microchannel. With these microdevices, excellent separation of the above analytes can be accomplished in relatively short times ( 2 minutes); with a limit of detection (LOD) in the sub-femto mole range. Different buffer systems were used including 2-[N-morpholino] ethanesulfonic acid (MES), phosphate, and borate buffers. Interestingly, significant improvement in the resolution was achieved by using borate buffer, where injecting a mixture of five analytes yield only two peaks with MES buffer, whereas five well-resolved peaks were obtained with borate buffer. These results show that these microdevices have promising separation resolving power and sensitivity for future medical diagnostics.
機(jī)譯：遺傳毒性致癌物可以與DNA共價(jià)反應(yīng)，形成DNA加合物，這可能導(dǎo)致關(guān)鍵基因發(fā)生突變，進(jìn)而導(dǎo)致癌癥等疾病的發(fā)展。因此，在涉及癌癥和其他衰老相關(guān)疾病的研究中，DNA損傷的作用及其隨后的生物標(biāo)記被認(rèn)為很重要。需要使用具有高分辨力和靈敏檢測(cè)能力的分析方法來檢測(cè)人體液體中的神經(jīng)遞質(zhì)（例如多巴胺（DA）和DA衍生的DNA加合物），兒茶酚雌激素衍生的DNA加合物和8-羥基-脫氧鳥苷DNA加合物。用紫外吸收檢測(cè)的標(biāo)準(zhǔn)色譜很難測(cè)定。此外，這些DNA加合物在室溫下熒光較弱。因此，天然熒光檢測(cè)方法對(duì)于此類生物標(biāo)志物無效。有趣的是，毛細(xì)管電泳（CE）與電化學(xué)檢測(cè)（ED）相結(jié)合是一種靈敏的選擇性技術(shù)，可用于分析包括這些分析物在內(nèi)的多種化合物。因此，制造了基于CE的PDMS /玻璃混合微流體裝置，用于具有完全集成的金電極的自由溶液電泳和簡(jiǎn)化的門控樣品注射，以分離和檢測(cè)各種DNA加合物和其他相關(guān)生物標(biāo)記。對(duì)ED系統(tǒng)的各種布置進(jìn)行了測(cè)試和評(píng)估，包括通道內(nèi)和通道內(nèi)檢測(cè)。最佳的電極配置是基于通道內(nèi)檢測(cè)進(jìn)行優(yōu)化的，該通道檢測(cè)采用原位制造的鈀解耦器作為位于微通道內(nèi)部的電泳接地。使用這些微型設(shè)備，可以在相對(duì)較短的時(shí)間內(nèi)（<2分鐘）完成上述分析物的出色分離。檢測(cè)限（LOD）處于亞毫微微摩爾范圍內(nèi)。使用了不同的緩沖液系統(tǒng)，包括2- [N-嗎啉代]乙磺酸（MES），磷酸鹽和硼酸鹽緩沖液。有趣的是，使用硼酸鹽緩沖液可實(shí)現(xiàn)分離度的顯著提高，其中注入五種分析物的混合物，使用MES緩沖液只能產(chǎn)生兩個(gè)峰，而使用硼酸鹽緩沖液則可以得到五個(gè)分辨良好的峰。這些結(jié)果表明，這些微型設(shè)備具有有前途的分離分辨能力和靈敏度，可用于未來的醫(yī)學(xué)診斷。

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