三种基因分型技术在病原微生物溯源方面的应用
发布时间：2018-10-12 21:40
【摘要】： 各种细菌性疾病一直危害着人类健康和畜牧业的健康发展。分型技术能够根据细菌的生化特征或基因组成分析不同来源菌株间的关系,从而对某次疾病暴发追踪溯源。过去的几十年里,大量基于DNA的基因分型技术发展起来,表现出传统分型技术无法比拟的优越性。基因分型技术要满足一些标准才能广泛推广,如分辨力、稳定性、简便快速、易于解释等。本研究主要比较脉冲场凝胶电泳PFGE、Sau-PCR和ERIC-PCR三种方法在微生物溯源中的应用,来探讨适于广泛推广的技术。 首先用三种方法对一起由福氏志贺菌引起的食物中毒事件进行溯源分析,分型结果高度一致,推论此次菌痢疫情的罪魁祸首是该餐馆中污染了福氏志贺菌的食物。据此建议相关食品监督部门和卫生检疫部门加强对其监测和管理。三种技术所用时间长短和成本花费多少的比较依次为PFGESau-PCRERIC-PCR。因此建议基层单位可首选便宜快速的ERIC-PCR或Sau-PCR进行初期的溯源分析。 其次利用三种方法对传代培养30代和室温放置30天的志贺菌进行基因分型分析,验证这三种方法带型的稳定性。PFGE带型未发生可检测的变化,具有完美的重复性。而ERIC-PCR发生了较大变化,容易提供错误的流行病学信息。发现Sau-PCR对于传代培养和室温放置的志贺菌分型结果都非常稳定,使实验室间的数据交流和建立数据库成为可能。进一步论证了Sau-PCR推广使用的可行性。 最后比较了Sau-PCR和ERIC-PCR技术对9株耐甲氧西林金黄色葡萄球菌(MRSA)的分型研究。结果显示MRSA的Sau-PCR基因型与SCCmec(staphylococcal cassette chromosome mec)基因型一致。而Sau-PCR技术无需了解菌株的基因信息,不需要特殊设备,具有操作简便快捷的优点,为MRSA的快速检测和溯源分析提供了有力的工具。 本论文依次比较了三种分型技术对病原微生物溯源的简便快捷性、稳定性和对革兰氏阳性菌MRSA的分辨力,发现了新发展的Sau-PCR技术比PFGE简便易行,比ERIC-PCR稳定、分辨力更强,为新方法Sau-PCR在基层单位的推广使用提供了进一步的实验和理论依据。
[Abstract]:A variety of bacterial diseases have been harmful to human health and the healthy development of animal husbandry. Typing technique can analyze the relationship between different strains according to the biochemical characteristics or gene composition of bacteria, so as to trace the source of a disease outbreak. In the past few decades, a large number of genotyping techniques based on DNA have been developed, showing unparalleled superiority compared with traditional genotyping techniques. Genotyping techniques must meet some criteria to be widely used, such as resolution, stability, simplicity and rapidity, easy to explain, and so on. In this study, the application of pulsed field gel electrophoresis (PFGE,Sau-PCR) and ERIC-PCR in microbial traceability was compared. Three methods were used to trace the food poisoning caused by Shigella flexneri, and the results were highly consistent. It was concluded that the culprit of the disease was the food contaminated with Shigella flexneri in the restaurant. Accordingly, the relevant food supervision departments and health and quarantine departments should strengthen their monitoring and management. The length of time and cost of the three technologies are compared in order of PFGESau-PCRERIC-PCR. Therefore, it is suggested that grassroots units should choose cheap and fast ERIC-PCR or Sau-PCR for initial traceability analysis. Then three methods were used to analyze the genotyping of Shigella after 30 generations of passage culture and 30 days at room temperature to verify the stability of the three methods. The PFGE band type was not detectable and had perfect reproducibility. However, ERIC-PCR has changed greatly and it is easy to provide wrong epidemiological information. It was found that Sau-PCR was very stable for subculture and room temperature storage of Shigella, which made it possible to exchange data between laboratories and establish databases. The feasibility of popularizing Sau-PCR is further demonstrated. Finally, Sau-PCR and ERIC-PCR techniques were compared in the typing of 9 strains of methicillin-resistant Staphylococcus aureus (MRSA). The results showed that the Sau-PCR genotype of MRSA was consistent with that of SCCmec (staphylococcal cassette chromosome mec) genotype. Sau-PCR technology does not need to know the genetic information of the strain and does not need special equipment. It has the advantages of simple and fast operation. It provides a powerful tool for rapid detection and traceability analysis of MRSA. In this paper, we have compared the convenience, stability and resolution of three typing techniques to trace pathogenic microorganisms to MRSA. It is found that the newly developed Sau-PCR technique is easier than PFGE, more stable than ERIC-PCR, and has stronger resolution. It provides further experimental and theoretical basis for the popularization and application of the new method Sau-PCR in basic units.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R37;S852.6
本文编号：2267694
[Abstract]:A variety of bacterial diseases have been harmful to human health and the healthy development of animal husbandry. Typing technique can analyze the relationship between different strains according to the biochemical characteristics or gene composition of bacteria, so as to trace the source of a disease outbreak. In the past few decades, a large number of genotyping techniques based on DNA have been developed, showing unparalleled superiority compared with traditional genotyping techniques. Genotyping techniques must meet some criteria to be widely used, such as resolution, stability, simplicity and rapidity, easy to explain, and so on. In this study, the application of pulsed field gel electrophoresis (PFGE,Sau-PCR) and ERIC-PCR in microbial traceability was compared. Three methods were used to trace the food poisoning caused by Shigella flexneri, and the results were highly consistent. It was concluded that the culprit of the disease was the food contaminated with Shigella flexneri in the restaurant. Accordingly, the relevant food supervision departments and health and quarantine departments should strengthen their monitoring and management. The length of time and cost of the three technologies are compared in order of PFGESau-PCRERIC-PCR. Therefore, it is suggested that grassroots units should choose cheap and fast ERIC-PCR or Sau-PCR for initial traceability analysis. Then three methods were used to analyze the genotyping of Shigella after 30 generations of passage culture and 30 days at room temperature to verify the stability of the three methods. The PFGE band type was not detectable and had perfect reproducibility. However, ERIC-PCR has changed greatly and it is easy to provide wrong epidemiological information. It was found that Sau-PCR was very stable for subculture and room temperature storage of Shigella, which made it possible to exchange data between laboratories and establish databases. The feasibility of popularizing Sau-PCR is further demonstrated. Finally, Sau-PCR and ERIC-PCR techniques were compared in the typing of 9 strains of methicillin-resistant Staphylococcus aureus (MRSA). The results showed that the Sau-PCR genotype of MRSA was consistent with that of SCCmec (staphylococcal cassette chromosome mec) genotype. Sau-PCR technology does not need to know the genetic information of the strain and does not need special equipment. It has the advantages of simple and fast operation. It provides a powerful tool for rapid detection and traceability analysis of MRSA. In this paper, we have compared the convenience, stability and resolution of three typing techniques to trace pathogenic microorganisms to MRSA. It is found that the newly developed Sau-PCR technique is easier than PFGE, more stable than ERIC-PCR, and has stronger resolution. It provides further experimental and theoretical basis for the popularization and application of the new method Sau-PCR in basic units.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R37;S852.6
【引证文献】
相关期刊论文 前1条
1 张淑红;吴清平;徐晓可;张菊梅;郭伟鹏;;Sau-PCR分子分型技术及应用[J];中国食品卫生杂志;2012年02期
，本文编号：2267694
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