三種基因分型技術(shù)在病原微生物溯源方面的應(yīng)用
發(fā)布時(shí)間：2018-10-12 21:40
【摘要】： 各種細(xì)菌性疾病一直危害著人類(lèi)健康和畜牧業(yè)的健康發(fā)展。分型技術(shù)能夠根據(jù)細(xì)菌的生化特征或基因組成分析不同來(lái)源菌株間的關(guān)系,從而對(duì)某次疾病暴發(fā)追蹤溯源。過(guò)去的幾十年里,大量基于DNA的基因分型技術(shù)發(fā)展起來(lái),表現(xiàn)出傳統(tǒng)分型技術(shù)無(wú)法比擬的優(yōu)越性�；蚍中图夹g(shù)要滿足一些標(biāo)準(zhǔn)才能廣泛推廣,如分辨力、穩(wěn)定性、簡(jiǎn)便快速、易于解釋等。本研究主要比較脈沖場(chǎng)凝膠電泳PFGE、Sau-PCR和ERIC-PCR三種方法在微生物溯源中的應(yīng)用,來(lái)探討適于廣泛推廣的技術(shù)。 首先用三種方法對(duì)一起由福氏志賀菌引起的食物中毒事件進(jìn)行溯源分析,分型結(jié)果高度一致,推論此次菌痢疫情的罪魁禍?zhǔn)资窃摬宛^中污染了福氏志賀菌的食物。據(jù)此建議相關(guān)食品監(jiān)督部門(mén)和衛(wèi)生檢疫部門(mén)加強(qiáng)對(duì)其監(jiān)測(cè)和管理。三種技術(shù)所用時(shí)間長(zhǎng)短和成本花費(fèi)多少的比較依次為PFGESau-PCRERIC-PCR。因此建議基層單位可首選便宜快速的ERIC-PCR或Sau-PCR進(jìn)行初期的溯源分析。 其次利用三種方法對(duì)傳代培養(yǎng)30代和室溫放置30天的志賀菌進(jìn)行基因分型分析,驗(yàn)證這三種方法帶型的穩(wěn)定性。PFGE帶型未發(fā)生可檢測(cè)的變化,具有完美的重復(fù)性。而ERIC-PCR發(fā)生了較大變化,容易提供錯(cuò)誤的流行病學(xué)信息。發(fā)現(xiàn)Sau-PCR對(duì)于傳代培養(yǎng)和室溫放置的志賀菌分型結(jié)果都非常穩(wěn)定,使實(shí)驗(yàn)室間的數(shù)據(jù)交流和建立數(shù)據(jù)庫(kù)成為可能。進(jìn)一步論證了Sau-PCR推廣使用的可行性。 最后比較了Sau-PCR和ERIC-PCR技術(shù)對(duì)9株耐甲氧西林金黃色葡萄球菌(MRSA)的分型研究。結(jié)果顯示MRSA的Sau-PCR基因型與SCCmec(staphylococcal cassette chromosome mec)基因型一致。而Sau-PCR技術(shù)無(wú)需了解菌株的基因信息,不需要特殊設(shè)備,具有操作簡(jiǎn)便快捷的優(yōu)點(diǎn),為MRSA的快速檢測(cè)和溯源分析提供了有力的工具。 本論文依次比較了三種分型技術(shù)對(duì)病原微生物溯源的簡(jiǎn)便快捷性、穩(wěn)定性和對(duì)革蘭氏陽(yáng)性菌MRSA的分辨力,發(fā)現(xiàn)了新發(fā)展的Sau-PCR技術(shù)比PFGE簡(jiǎn)便易行,比ERIC-PCR穩(wěn)定、分辨力更強(qiáng),為新方法Sau-PCR在基層單位的推廣使用提供了進(jìn)一步的實(shí)驗(yàn)和理論依據(jù)。
[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.
【學(xué)位授予單位】：華南理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R37;S852.6
本文編號(hào)：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.
【學(xué)位授予單位】：華南理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R37;S852.6
【引證文獻(xiàn)】
相關(guān)期刊論文 前1條
1 張淑紅;吳清平;徐曉可;張菊梅;郭偉鵬;;Sau-PCR分子分型技術(shù)及應(yīng)用[J];中國(guó)食品衛(wèi)生雜志;2012年02期
，本文編號(hào)：2267694
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