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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 周崇熙 | |
dc.contributor.author | Chung-Wen Hu | en |
dc.contributor.author | 胡崇玟 | zh_TW |
dc.date.accessioned | 2021-05-15T18:00:20Z | - |
dc.date.available | 2015-02-26 | |
dc.date.available | 2021-05-15T18:00:20Z | - |
dc.date.copyright | 2014-02-26 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-01-17 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5485 | - |
dc.description.abstract | Salmonella enterica serovar Schwarzengrund 是一具有人畜共通傳染風險之血清型,相較於歐美國家,臺灣於家禽、豬隻、人類都曾經分離出高比例的S. Schwarzengrund,但是在歐美國家S. Schwarzengrund卻鮮少被分離,非主要造成動物或人類疾病的血清型。本研究利用30株於2000-2012年來自鴨、寵物食品、雞、野鳥、豬、火雞等不同來源之S. Schwarzengrund菌株,因具有流行病學相關性的菌株會有相同的抗藥性表現,因此本研究中分析30株S. Schwarzengrund 對於Ampicillin、Cephalothin、Chloramphenicol、Gentamicin、Streptomycin、Kanamycin、Tetracycline、Oxytetracycline、Ciprofloxacin、Enrofloxacin、Nalidixic acid以及Sulfamethoxazole/ Trimethoprim等12種抗菌劑的抗藥性差異,利用抗藥性表現以及流行病學背景篩選出不具流行病學相關性的菌株,用來計算分子分型法鑑別力時。進行脈衝式電泳分析時,發現常用的XbaI限制酶分型法無法有效辨別出無流行病學相關性之菌株,進而使用AvrII、SpeI、NotI、SfiI、PacI等五種限制酶,觀察其對於 S. Schwarzengrund 的分型能力。並且進一步利用Multi Locus Sequence Typing (MLST)及介質輔助雷射脫附飛行時間質譜法(MALDI-TOF MS),評估這些方法的分型鑑別能力。脈衝式電泳的結果,對於不具流行病學相關之S. Schwarzengrund菌株XbaI限制酶D值為0.65,AvrII、SpeI、NotI、SfiI、PacI等限制酶之D值分別為0.96、0.89、0.95、0.93、0.91。使用AvrII以及SfiI對於S. Schwarzengrund菌株之分型力較佳,若結合此兩個限制酶,可以使得不具流行病學相關性的菌株被成功的鑑別,推薦可使用在S. Schwarzengrund的分型。本試驗中MLST使用aroC、dnaN、hemD、hisD、purE、sucA、thrA等七個管家基因作為定序的目標,若有單一點突變即可予以辨別,由序列比對得知各菌株之sequence type。MLST結果 S. Albany、S. Enteritidis、S. Typhimurium、S. Schwarzengrund以及S. Pullorum之sequence type分別為ST292、ST11、ST19、ST96、ST92,可見MLST能夠成功的鑑別不同血清型,但30株S. Schwarzengrund皆為ST96,因此MLST對於S. Schwarzengrund不具分型能力。利用MALDI-TOF MS雖可以將30株S. Schwarzengrund菌株分為A1、A2、B、C等4個群集,但同一來源之S. Schwarzengrund卻被鑑定為不同之群集,且MALDI-TOF MS也無法將不同血清型做區別,因此MALDI-TOF並不適合作為S. Schwarzengrund的親緣性鑑定。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-05-15T18:00:20Z (GMT). No. of bitstreams: 1 ntu-103-R00629028-1.pdf: 3139237 bytes, checksum: 9c4d76937a85df1bf80343ec3278a0b8 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 致謝 I
摘要 III Abstract V 目錄 VII 表次 IX 圖次 X 第一章 緒言 1 第二章 文獻探討 3 第一節 沙門氏菌簡介 3 2.1.1. 沙門氏菌特性 3 2.1.3. 沙門氏菌之流行病學 5 2.1.4. 沙門氏菌之抗藥性 7 第二節 Schwarzengrund 血清型沙門氏菌 12 2.2.1. S. Schwarzengrund簡介 12 2.2.2. S. Schwarzengrund 流行病學 13 第三節 分子生物學分型方法 16 2.3.1. 脈衝式電泳 16 2.3.2. 多重基因座序列分型法 (MLST) 19 2.3.3. 介質輔助雷射脫附-飛行時間質譜法 (MALDI-TOF) 21 2.3.4. 分子分型法之鑑別力 26 第三章 材料與方法 27 第一節 沙門氏菌來源及鑑定 27 3.1.1. Schwarzengrund血清型沙門氏菌來源 27 3.1.2. 沙門氏菌血清型之鑑定 27 第二節 沙門氏菌之抗菌劑感受性試驗 30 3.2.1 培養基配置 30 3.2.2 菌液濃度調整 30 3.2.3 瓊脂藥片擴散法 30 第三節 脈衝式電泳 32 3.3.1 試劑製備 32 3.3.2 菌塊製備 32 3.3.3 限制酶切割 33 3.3.4 PFGE結果 34 第四節 多重基因座序列分型法 35 3.4.1. 萃取Genomic DNA 35 3.4.2. 基因片段增幅及定序 35 第五節 介質輔助雷射脫附飛行時間質譜法 37 第四章 結果 38 第一節 抗菌劑感受性試驗 38 第二節 脈衝式電泳 39 第三節 多重基因座序列分型法 45 第四節 介質輔助雷射脫附飛行時間質譜法(MALDI-TOF) 47 第五章 討論 48 第一節 抗菌劑感受性試驗 48 第二節 脈衝式電泳 52 第三節 多重基因座序列分型法 56 第四節 介質輔助雷射脫附飛行時間質譜法 59 第六章 結論與建議 62 第七章 參考文獻 64 表格 72 圖片 80 | |
dc.language.iso | zh-TW | |
dc.title | Schwarzengrund 血清型沙門氏菌之分子分型法之比較 | zh_TW |
dc.title | Comparison of Molecular Subtyping Methods for Salmonella enterica serovar Schwarzengrund | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-1 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 林辰栖 | |
dc.contributor.oralexamcommittee | 連一洋 | |
dc.subject.keyword | Schwarzengrund血清型沙門氏菌,脈衝式電泳,多重基因座序列分型法,介質輔助雷射脫附飛行時間質譜法, | zh_TW |
dc.subject.keyword | Salmonella enterica serovar Schwarzengrund,Pulsed field gel electrophoresis,Multi locus sequence typing,Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry, | en |
dc.relation.page | 101 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2014-01-20 | |
dc.contributor.author-college | 獸醫專業學院 | zh_TW |
dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
顯示於系所單位: | 獸醫學系 |
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