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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊瑋誠(Wei-Cheng Yang) | |
| dc.contributor.author | Meng-Jung Hsieh | en |
| dc.contributor.author | 謝孟容 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:06:35Z | - |
| dc.date.available | 2022-01-17 | |
| dc.date.available | 2022-11-24T03:06:35Z | - |
| dc.date.copyright | 2022-01-17 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-12-29 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80433 | - |
| dc.description.abstract | "弓蟲病是一個重要的人畜共通傳染病，不管是在獸醫學以及公共衛生上都十分重視。在鯨豚身上發現陸地上的弓蟲病，可以視為是一個從陸地到海洋的病源污染。目前，弓蟲診斷需要在專業的實驗室裡進行，因此弓蟲的病例報告主要來自某些特定的國家。此外，當擱淺事件發生在含有許多島嶼的熱帶國家，並不是每個小島都有後送的實驗室。由於交通運輸不便，運送樣本時間過長，加上天氣炎熱，以至於擱淺動物樣本難以良好保存，使得DNA容易降解讓疾病監測更加困難。為了監測全球的擱淺鯨豚的弓蟲感染情形，我們研發可攜帶性之隔絕式恆溫PCR (insulated isothermal PCR, iiPCR)，搭配小型自動磁珠萃取，達到野外檢測之目的。我們使用B1基因當作檢測弓蟲的目標基因，同時使用鯨豚的β2-microglobulin (B2M)基因當作內部控制。將弓蟲質體加入鯨豚大腦、小腦、心肌、肌肉模擬臨床樣本，進行靈敏度分析。結果顯示B1/B2M (B1 gene/ β2-microglobulin) iiPCR在25 mg的組織下，只要21~86個緩殖體就可以檢測出陽性，B1/B2M iiPCR與qPCR有幾乎完美一致性(92%; 95% CI: 0.78–0.90; κ=0.84)。雙重B1/B2M iiPCR檢測平台不只方便攜帶去野外檢測，且敏感性高也更快速得到檢驗結果，適合運用在擱淺鯨豚之弓蟲檢驗。此平台提供一個有效方法，以鯨豚作為哨兵動物，評估並監測目前海洋環境生態的健康。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:06:35Z (GMT). No. of bitstreams: 1 U0001-2212202113115100.pdf: 4334215 bytes, checksum: 7bb5287c4a43ca195d558debba67e3f9 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "中文摘要 I ABSTRACT II SECTION 1 INTRODUCTION 1 1.1 Toxoplasma gondii 1 1.1.1 History 1 1.1.2 Life cycle 2 1.2 Transmission and host 4 1.2.1 Transmission 4 1.2.2 Definitive host 5 1.2.3 Intermediate host 5 1.2.4 Human 7 1.3 Transmission in water 8 1.3.1 Marine mammals 11 1.4 Diagnosis 16 1.4.1 Traditional, non-DNA-based diagnostic methods 16 1.4.2 Molecular assays 17 1.4.3 Genotyping 20 1.5 Research goal 22 SECTION 2 MATERIALS AND METHODS 24 2.1 Ethic statement 24 2.2 Synthetic spike-in standard 24 2.3 DNA extraction 26 2.4 Insulated Isothermal PCR 26 2.5 Real-time PCR 29 2.6 Statistics 29 SECTION 3 RESULTS 30 3.1 DNA concentration 30 3.2 Analytical specificity 31 3.3 Analytical sensitivity 32 3.4 Sensitivity in singleplex iiPCR 33 3.5 Sensitivity in duplex iiPCR 34 3.6 Performance evaluation of the B1/B2M iiPCR 35 3.7 Preliminary result in clinical samples 39 SECTION 4 DISCUSSION 40 SECTION 5 REFERENCE 45 Figures： Fig. 1 Enteroepithelial stages of T. gondii in epithelial cells of the small intestine from a domestic cat. 3 Fig. 2 Major routes of transmission of T. gondii. 4 Fig. 3 A route of T. gondii oocyst-induced transmission. 10 Fig. 4 The asexual stage of T. gondii in cetaceans after ingesting oocysts or tissue cysts. 12 Fig. 5 The world map of T. gondii or T. gondii–like protozoan parasite exposure in cetaceans. 14 Fig. 6 POCKIT™ system workflow for point-of-need detection of T. gondii synthetic spike-in standard. 23 Fig. 7 A: The map of synthetic spike-in standard; B: The sequence of synthetic spike-in standard. 25 Fig. 8 The boxplot of concentration of DNA. 30 Tables： Table 1 T. gondii or T. gondii–like protozoan parasite exposure in cetaceans. 15 Table 2 The sequence of primers and probes. 28 Table 3 Analytical sensitivity of singleplex iiPCR. 32 Table 4 Sensitivity in sigleplex iiPCR. 33 Table 5 Sensitivity in duplex iiPCR. 34 Table 6 Performance evaluation of the B1/B2M iiPCR using T. gondii-spiked cetacean samples. 38 Table 7 Preliminary result in twenty-five tissue samples of six individual. 39 Table 8 Preliminary result in twenty-five tissue samples from different organs. 39 " | |
| dc.language.iso | en | |
| dc.subject | 污染 | zh_TW |
| dc.subject | 可攜帶式 | zh_TW |
| dc.subject | 隔絕式恆溫聚合酶連鎖反應 | zh_TW |
| dc.subject | 鯨豚 | zh_TW |
| dc.subject | 弓蟲 | zh_TW |
| dc.subject | Toxoplasma | en |
| dc.subject | pollution | en |
| dc.subject | insulated isothermal PCR | en |
| dc.subject | cetaceans | en |
| dc.subject | field-deployed | en |
| dc.title | 可攜帶型隔絕式恆溫聚合酶連鎖反應檢測鯨豚弓蟲感染之研發 | zh_TW |
| dc.title | Development of a field-deployable insulated isothermal PCR (iiPCR) for the detection of Toxoplasma gondii in cetaceans | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 詹昆衛(Hsin-Tsai Liu),王浩文(Chih-Yang Tseng) | |
| dc.subject.keyword | 可攜帶式,隔絕式恆溫聚合酶連鎖反應,鯨豚,弓蟲,污染, | zh_TW |
| dc.subject.keyword | insulated isothermal PCR,cetaceans,Toxoplasma,pollution,field-deployed, | en |
| dc.relation.page | 73 | |
| dc.identifier.doi | 10.6342/NTU202104557 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-12-30 | |
| 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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| U0001-2212202113115100.pdf 授權僅限NTU校內IP使用（校園外請利用VPN校外連線服務） | 4.23 MB | Adobe PDF |
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