葡萄球菌属的鉴定及分类方法研究

发布时间：2018-04-27 01:25

本文选题：葡萄球菌 + 金黄色葡萄球菌　；参考：《贵州大学》2006年硕士论文

【摘要】：随着食物中毒事件的频繁发生，食品安全问题显得越来越重要。HACCP就是为了控制食品安全卫生而发展起来的一种国际食品安全控制体系，它是英文Hazard Analysis and Critical Control Point的缩写，即“危害分析与关键控制点”系统。HACCP是人们用来控制食品安全危害的一种通常技术，一个分析工具，也是一种重要的管理体系，它可以与任何操作相结合，能使正在进行的食品安全性项目保持经济有效。葡萄球菌属在自然界中分布广泛，空气、水、灰尘及人和动物的排泄物中都可以找到。本属中金黄色葡萄球菌(金葡菌)是一种重要的食源性致病菌，为革兰氏阳性球状菌，直径0.8-1.0μm，呈葡萄状排列，有些菌株还具有荚膜或粘层。金葡菌是引起食品污染和细菌性食物中毒的一种主要细菌，食品受其污染后，不仅腐败变质，而且部分菌株产生肠毒素，肠毒素是引起食物中毒的主要原因。目前研究发现葡萄球菌属的其他部分种也成为人们关注的致病菌。我国已加入WTO，，这对我国的食品加工行业来说，既是机遇也是挑战。为了使我们的食品更顺利地走向国际市场，有必要对金葡菌及葡萄球菌属的其它种可能导致的危害进行准确地分析。本文主要运用经典方法及分子生物学方法对葡萄球菌进行研究，旨在为葡萄球菌的分类及葡萄球菌检测试剂盒的研制方法提供依据。本文从3个部分对葡萄球菌属进行研究： 1．葡萄球菌属的经典方法鉴定及肠毒素测定该部分主要是收集供试菌株，然后对其进行经典方法进行鉴定和肠毒素的测定。试验主要运用革兰氏染色及API Staph系统进行菌种的鉴定；酶联免疫法进行肠毒素的测定。结果表明：供试的15株菌都为葡萄球菌；在分析中API Staph系统存在的弊端是当颜色反应不完全时，对结果判断的影响较大；葡萄球菌属中不仅重要致病菌金葡菌产生肠毒素，其他一些葡萄球菌也产生肠毒素，其中1，2，3，6，7，13号菌株为产毒素菌株，其余菌株为不产毒素菌株；因此，以往研究较少的非致病菌也应该成为食品检测关注的对象。 2．葡萄球菌属菌株rDNA的ITS序列分析本文对15株葡萄球菌的ITS序列进行扩增，得到10株菌的序列，并对10株菌及其GenBank中的相似序列进行比对及系统树的构建。结果显示：ITS序列能为葡萄球菌属的分类及检测试剂盒的研制提供较好的指导作用。
[Abstract]:With the frequent occurrence of food poisoning, food safety becomes more and more important. HACCP is an international food safety control system developed to control food safety and hygiene. It is the Hazard Analysis and Critical The acronym of Control Point, or "Hazard Analysis and critical Control Point" system, is a common technology, an analytical tool, and an important management system that people use to control food safety hazards, and it can be combined with any operation. To keep ongoing food safety programs economical and effective. Staphylococcus is widely distributed in nature and can be found in air, water, dust, and human and animal excreta. Staphylococcus aureus (Staphylococcus aureus) is an important foodborne pathogenic bacteria. It is a Gram-positive globular bacteria with diameter of 0.8-1.0 渭 m and grape-like arrangement. Some strains also have capsule or mucilage. Staphylococcus aureus is one of the main bacteria that cause food contamination and bacterial food poisoning. Food contaminated by Staphylococcus aureus not only spoils and spoils, but also produces enterotoxin, which is the main cause of food poisoning. Other species of the genus Staphylococcus have also been found to be the cause of concern. China has joined WTO, which is both an opportunity and a challenge for China's food processing industry. In order to make our food more smoothly to the international market, it is necessary to accurately analyze the possible harm caused by Staphylococcus aureus and other species of Staphylococcus. In this paper, the classical method and molecular biological method were used to study Staphylococcus in order to provide the basis for the classification of Staphylococcus and the development of staphylococcal detection kit. In this paper, the genus Staphylococcus was studied in three parts: 1. Identification of Staphylococcus and determination of Enterotoxin this part is mainly to collect the strains tested and then to identify the classical method and the determination of enterotoxin. Gram staining and API Staph system were used to identify the bacteria, and enzyme linked immunosorbent assay (Elisa) was used to detect enterotoxin. The results showed that all the 15 strains were staphylococci, the disadvantage of API Staph system in analysis was that when the color reaction was not complete, it had a great influence on the result judgment, Staphylococcus not only produced enterotoxin, but also produced enterotoxin in Staphylococcus, which was an important pathogen in the genus Staphylococcus. Some other staphylococci also produce enterotoxin, among them, strain No. 13 is a toxin producing strain, while the others are non-toxin producing strains. Therefore, the non-pathogenic bacteria which have been studied in the past should also be the focus of food detection. 2. ITS sequence Analysis of Staphylococcus strain rDNA in this paper, the ITS sequences of 15 strains of Staphylococcus were amplified, and the sequences of 10 strains of Staphylococcus were obtained. The similar sequences of 10 strains and their GenBank were compared and the phylogenetic tree was constructed. The results showed that its sequence could provide good guidance for the classification of Staphylococcus genus and the development of detection kit.
【学位授予单位】：贵州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378.1;Q939

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