本文選題：大腸桿菌 + 富集��； 參考：《大連理工大學》2016年碩士論文

【摘要】：關注共享水的安全問題,是一個十分重要的社會問題。由于大腸桿菌感染劑量很低,并且是許多食源性和水源性胃腸炎的發(fā)病源頭,因此對于大腸桿菌的檢測十分重要。為了能夠準確檢測水中的大腸桿菌,人們一直致力于發(fā)展一些具有生物特異性的方法。而多種技術(shù)的開發(fā),已被證明難以實現(xiàn)所有相關類型病毒的檢測。同時少量采樣和局部采樣,對實際的生活和生產(chǎn)中的大量水樣起不到有效的檢測。因而大量地,快速地富集水樣對于檢測就成為一個關鍵性的手段。本文討論了一些常用的富集手段,并且綜合前人的經(jīng)驗和努力,致力于發(fā)展一種新的手段和方法來提高細菌的富集效率,縮短富集的時間,為傳統(tǒng)的細菌培養(yǎng)技術(shù)和新型的生物分子監(jiān)測技術(shù)提供支持,提高腸道病菌的檢測效率和準確性。本論文當中采用兩步富集方式對水體中的大腸桿菌0157：H7進行富集回收和檢測。一步富集方式即超濾,使用聚丙烯膜對0157：H7進行富集回收,并且在超濾過程中采取反沖洗方式,提高了一步富集的回收效率,富集效率最高可達67.21%；二步富集方式中采用磁性包被粒子對0157：H7進行回收。包被物質(zhì)均采用無毒,并且生物相容性好的一些物質(zhì),合成了氨基磁珠,羧基磁珠,和殼聚糖包被磁珠,對其磁性,XRD,紅外等性質(zhì)作了表征,并對其等電點進行了測定。比較了三種磁珠不同pH值,不同磁珠量,不同吸附時間下的吸附效果。發(fā)現(xiàn)氨基磁珠磁珠量在2.5ml時,對大腸桿菌的吸附能力可以達到百分之百,提高了對大腸桿菌的二步富集效率。通過兩步富集手段,將水體的體積濃縮至可以測定的范圍,濃縮倍數(shù)達到500倍。降低了由大體積水樣的局部取樣造成的測定結(jié)果不準確的概率,為細菌檢測提供了保障。
[Abstract]:It is a very important social problem to pay attention to the safety of shared water. The detection of Escherichia coli is very important because of the low dose of Escherichia coli and the fact that it is the source of many gastroenteritis caused by food and water. In order to detect E. coli in water accurately, some biological specific methods have been developed. However, the development of various technologies has proved difficult to detect all the related viruses. At the same time, a small amount of sampling and local sampling can not effectively detect a large number of water samples in real life and production. Therefore, a large number of rapid enrichment of water samples for detection becomes a key means. In this paper, some common enrichment methods are discussed, and based on previous experiences and efforts, a new method is developed to improve the efficiency of bacteria enrichment and shorten the time of enrichment. It provides support for traditional bacterial culture technology and new biomolecular monitoring technology to improve the detection efficiency and accuracy of intestinal bacteria. In this paper, two-step enrichment method was used for the recovery and detection of Escherichia coli 0157:H7 in water. In the process of one step enrichment, ultrafiltration is used to enrich and recover 0157:H7 with polypropylene membrane, and backwashing is adopted in the process of ultrafiltration, which improves the recovery efficiency of one step enrichment. The highest enrichment efficiency is 67.21, and the magnetic encapsulated particles are used to recover 0157:H7 in the two-step enrichment. Aminomagnetic beads, carboxyl beads and chitosan coated beads were synthesized by using nontoxic and biocompatible materials. The magnetic properties of the beads were characterized by XRDand IR, and their isoelectric points were determined. The adsorption effects of three kinds of magnetic beads under different pH value, different amount of magnetic beads and different adsorption time were compared. It was found that when the amount of magnetic beads of amino magnetic beads was 2.5ml, the adsorption ability of Escherichia coli could reach 100%, which improved the efficiency of two-step enrichment of Escherichia coli. By means of two-step enrichment, the volume of water was condensed to the measurable range, and the concentration multiple reached 500 times. The probability of inaccuracy caused by local sampling of large water samples is reduced, which provides a guarantee for bacterial detection.
【學位授予單位】：大連理工大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R123.1

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