本文關(guān)鍵詞：基于熒光標記發(fā)夾式DNA探針的汞離子定量檢測技術(shù)的研究　出處：《太原理工大學》2017年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 汞污染 DNA鏈 熒光染料 熒光能量共振轉(zhuǎn)移 定量檢測

【摘要】：環(huán)境污染已成為一個全球性問題,而重金屬污染在環(huán)境污染中備受關(guān)注,汞污染作為較嚴重的一種重金屬污染,對生態(tài)環(huán)境具有嚴重的危害。由于汞難被降解,能夠在土壤、水、空氣中廣泛富集,最后經(jīng)過食物鏈和大氣等進入人體,嚴重破壞蛋白和酶的活性,造成器官中毒。為了更好的預防汞污染帶來的食品安全、疾病危機等問題,開發(fā)一種方便快捷、靈敏高效,并且能夠?qū)崿F(xiàn)現(xiàn)場檢測,用于臨床診斷的檢測手段變得越來越重要。在汞離子的檢測方法中使用最廣泛的屬于常規(guī)的大型儀器檢測方法,如原子吸收光譜法、ICP-MS等方法,這些方法在檢測靈敏度和精度上都非常高,但是操作比較復雜,而且耗時也相對較長。而面對日益嚴峻復雜的汞污染挑戰(zhàn),為了實現(xiàn)對不同環(huán)境現(xiàn)場的汞污染檢測,研究人員也在不斷開發(fā)一些新型的Hg~(2+)檢測方法。如試紙條法、化學傳感器法,納米材料傳感器法以及DNA探針法等,試紙條法是比較簡便的一種方法,但只能做定性檢測,無法實現(xiàn)定量分析,而其他檢測方法基本都是利用化學分子或一些官能團能夠結(jié)合Hg~(2+),引起分子結(jié)構(gòu)或者分子的發(fā)光特性的改變,最后通過觀察吸收譜或光譜來實現(xiàn)對Hg~(2+)的定量檢測,相對傳統(tǒng)大型儀器方法,這些方法在操作和成本上具有很大改善,但在靈敏度和耗時上仍需改進。結(jié)合DNA鏈特性和熒光共振能量轉(zhuǎn)移技術(shù),在本課題中我們設(shè)計了一種發(fā)夾式的富含T堿基的DNA鏈,無需繁瑣的溫度處理,實現(xiàn)在常溫下一步法檢測Hg~(2+)。在DNA鏈兩端分別修飾熒光染料Cy3、Cy5,通過觀察Cy3熒光強度的變化,將其變化與Hg~(2+)濃度進行線性擬合,我們發(fā)現(xiàn)隨著Hg~(2+)濃度的增加,Cy3的熒光強度在不斷增大,它們之間具有線性相關(guān)性,并且線性相關(guān)系數(shù)為0.9836,而且我們的方法最低檢測限能夠達到1.3 nM,低于國家飲用水中Hg~(2+)的標準檢測限5 nM。隨后對方法的選擇性進行了研究,加入不同的干擾離子,發(fā)現(xiàn)我們的方法對Hg~(2+)具有好的選擇性,能夠特異性識別Hg~(2+),實現(xiàn)了對Hg~(2+)的選擇性的定量檢測。
[Abstract]:Environmental pollution has become a global problem, and heavy metal pollution in the environmental pollution has received much attention. As a serious heavy metal pollution, mercury pollution has serious harm to the ecological environment, because mercury is difficult to be degraded. Can be widely enriched in soil, water, air, and finally through the food chain and atmosphere into the human body, seriously damage the activity of proteins and enzymes, resulting in organ poisoning. In order to better prevent mercury pollution brought food safety. Disease crisis and other problems, the development of a convenient, rapid, sensitive and efficient, and can achieve on-site detection. The detection methods for clinical diagnosis are becoming more and more important. The most widely used methods of mercury ion detection are the conventional large-scale instrument methods, such as atomic absorption spectrometry (AAS) and ICP-MS. These methods are very sensitive and accurate, but the operation is complex and time-consuming, and face the increasingly serious challenges of mercury pollution. In order to detect mercury pollution in different environmental sites, researchers have been developing some new Hg~(2 detection methods, such as test strip method, chemical sensor method. Nanomaterial sensor method and DNA probe method, the test strip method is a relatively simple method, but can only do qualitative testing, can not achieve quantitative analysis. Other detection methods are based on the use of chemical molecules or some functional groups can bind to Hg~(2, resulting in changes in molecular structure or molecular luminescence properties. Finally, the quantitative detection of Hg~(2 is realized by observing the absorption spectrum or spectrum. Compared with the traditional large-scale instrument method, these methods have a great improvement in operation and cost. However, we still need to improve the sensitivity and time consuming. Combining with the characteristics of DNA chain and fluorescence resonance energy transfer technology, we designed a hairpin DNA chain rich in T-base. The Hg~(2 can be detected by one step method at room temperature without complicated temperature treatment. The fluorescent dye Cy3C Cy5 is modified at the two ends of DNA chain, and the change of fluorescence intensity of Cy3 is observed. By linear fitting with the concentration of Hg~(2, we found that the fluorescence intensity of Cy3 increased with the increase of the concentration of Hg~(2, and there was a linear correlation between them. The linear correlation coefficient is 0.9836, and the minimum detection limit of our method can reach 1.3 nm. The detection limit of 5 nm was lower than the standard detection limit of Hg~(2 in drinking water. Then the selectivity of the method was studied and different interference ions were added. It is found that our method has good selectivity for Hg~(2, and can specifically identify Hg~(2, thus realizing the selective quantitative detection of Hg~(2.
【學位授予單位】：太原理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O657.3

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