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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 葉信宏(Hsin-Hung Yeh) | |
dc.contributor.author | Pei-Ling Huang | en |
dc.contributor.author | 黃佩玲 | zh_TW |
dc.date.accessioned | 2021-06-08T03:49:29Z | - |
dc.date.copyright | 2018-12-17 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-12-06 | |
dc.identifier.citation | 1.林于勤、李淑英、陳豪勇、林鼎翔。2014。新型恆溫式圈環形核酸增幅法Loop-mediated Isothermal Amplification (LAMP) 簡介與應用。疫情報導 20 (6):323-332。
2.吳榮彬。2010。台灣梨樹病毒病害之鑑定及發展應用於田間調查和檢疫之檢測技術。國立中興大學植物病理學系博士論文。174 pp。 3.吳榮彬、詹富智。2007。梨輪紋嵌紋病、梨脈黃化病、梨莖凹陷病。P.75-89。梨樹保護。臺北市:行政院農業委員會動植物防疫檢疫局。155 pp。 4.林嘉興、廖萬正、林信山、林長仁。1991。梨栽培之回顧與展望。P.379-396。臺灣果樹之生產及研究發展研討會專刊。 5.黃振文、蔡東纂、曾國欽、詹富智。2001。梨樹病害圖鑑。臺中市:行政院農業委員會動植物防疫檢疫局。57 pp。 6.張清安。1996。植物病毒鑑定及診斷新技術。植物保護新科技研討會專刊。P.35-45。臺灣省農業試驗所。 7.張雙納、李正男、范旭東、張尊平、任芳、胡國君、董雅鳳。2018。蘋果褪綠葉斑病毒 RT-LAMP 檢測方法的建立。中國農業科學 51 (9):1706-1716。 8.詹富智、吳榮彬、郭璦臻、鄭尤琇、張賀雄、蘇秋竹、楊耀祥。2003a。應用反轉錄聚合酶連鎖反應偵測蘋果黃化葉斑病毒和蘋果莖凹陷病毒。植物病理學會刊12 (1):12-18。 9.詹富智、盧耀村、陳慶忠。2003b。植物檢疫病毒偵測技術。P.35-52。植物重要防疫檢疫病害診斷鑑定技術研習會專刊(二)。臺北市:行政院農業委員會動植物防疫檢疫局。 10.劉秀玲、劉淑玲、林長平。2007。臺灣梨衰弱病。P.70-74。梨樹保護。臺北市:行政院農業委員會動植物防疫檢疫局。155 pp。 11.廖萬正。1987。高接梨接穗新梢之再利用。P.175-183。園藝作物產期調節研討會專集-特刊第10號。 12.廖萬正。2005a。梨。P.203-208。行政院農業委員會台灣農家要覽增修訂三版策劃委員會(編著)。臺灣農家要覽-農作篇(二)。臺北市:財團法人豐年社。 13.廖萬正。2005b。臺灣梨栽培技術之發展。P.47-54。梨栽培管理技術研討會專輯。 14.蘇秋竹。2007。梨葉緣焦枯病。P.64-69。梨樹保護。臺北市:行政院農業委員會動植物防疫檢疫局。155 pp。 15.中國大陸梨接穗有害生物風險評估Pathway-initiated Pest Risk Assessments for Pear Scion from Mainland China。2012。(防檢局內部資料未公開) 16.Aboubakr, M., Mohammadamin, A., Hamid, A., and Jaber, N. 2012. Development and evaluation of a loop-mediated isothermal amplification assay for detection of Erwinia amylovora based on chromosomal DNA. Eur. J. Plant Pathol. 133 (3): 609-620. 17.Adams, M. J., Antoniw, J. F., Bar-Joseph, M., Brunt, A. A., Candresse, T., Foster, G. D., Martelli, G. P., Milne, R. G., and Fauquet, C. M. 2004. The new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Arch. Virol. 149: 1045-1060. 18.Almasi, M. A., Karami, S., Nasiri, M., Moradi, A., and Nasiri, J. 2012. Assessment of performance ability of three diagnostic methods for detection of Potato leafroll virus (PLRV) using different visualizing systems. Appl. Biochem. Biotechnol. 168 (4): 770-784. 19.Beata, H. J., and Natasza, B. 2013. Detection of Pepino mosaic virus isolates from tomato by one-step reverse transcription loop-mediated isothermal amplification. Arch. Virol. 158 (10): 2153-2156. 20.Boubourakas, I. N., Fucuta, S., Luigi, M., Faggioli, F., Barba, M., and Kyriakopoulou, P. E. 2010. Improvement of the reverse transcription loop mediated isothermal amplification (RT-LAMP) method for the detection of Peach latent mosaic viroid (PLMVd). 2010. 21st International Conference on Virus and other Graft Transmissible Diseases of Fruit Crops. Julius-Kühn-Archiv 427: 65-69. 21.Chen, S., Zhou, Y., Ye, T., Hao, L., Guo, L., Fan, Z., Li, S., and Zhou, T. 2014. Genetic variation analysis of Apple chlorotic leaf spot virus coat protein reveals a new phylogenetic type and two recombinants in China. Arch. Virol. 159 (6): 1431-1438. 22.Fukuta, S., Iida, T., Mizukami, Y., Ishida, A., Ueda, J., Kanbe, M., and Ishimoto, Y. 2003. Detection of Japanese yam mosaic virus by RT-LAMP. Arch. Virol. 148: 1713-1720. 23.Fukuta, S., Ohishi, K., Yoshida, K., Mizukami, Y., Ishida, A., and Kanbe, M. 2004. Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of Tomato spotted wilt virus from chrysanthemum. J. Virol. Methods 121: 49-55. 24.Huang, C., Sun, Z. Y., Yan, J. H., Luo, Y., Wang, H. G., and Ma, Z. H. 2011. Rapid and precise detection of latent infections of wheat stripe rust in wheat leaves using loop-mediated isothermal amplification. J. Phytopathol. 159: 582-584. 25.Jones, A. L., and Aldwinckle, H. S. 1990. Compendium of Apple and Pear Diseases. American Phytopathological Society, Minnesota, USA, 100 pp. 26.Kanetani, S., Kikuchi, T., Akiba, M., Nakamura, K., Ikegame, H., and Tetsuka, K. 2011. Detection of Bursaphelenchus xylophilus from old discs of dead Pinus armandii var. amamiana trees using a new detection kit. For. Pathol. 41: 387-391. 27.Kikuchi, T., Aikawa, T., Oeda, Y., Karim, N., and Kanzaki, N. 2009. A rapid and precise diagnostic method for detecting the pinewood nematode Bursaphelenchus xylophilus by loop-mediated isothermal amplification. Phytopathology 99: 1365-1369. 28.Kim, S. H., Koh, G. C., Hwang, B., Lee, H., and Hong, S. W. 2009. Identification of diffenential gene expression in juvenile vs. mature leaves of pear (Pyrus pyrifolia) by using annealing control primer. J. Korean Soc. Appl. Biol. Chem. 52 (2): 121-127. 29.Kubota, R., Jenkins, D. M., Alvarez, A. M., and Vine, B. G. 2008. Detection of Ralstonia solanacearum by loop-mediated isothermal amplification. Phytopathology 98 (9): 1045-1051. 30.Kubota, R., Schell, M. A., Peckham, G. D., Rue, J., Alvarez, A. M., Allen, C., and Jenkins, D. M. 2011. In silico genomic subtraction guides development of highly accurate, DNA-based diagnostics for Ralstonia solanacearum race 3 biovar 2 and blood disease bacterium. J. Gen. Plant Pathol. 77: 182-193. 31.Li, X., Nie, J., Ward, L., Madani, M., Hsiang, T., Zhao, Y., and De Boer, S. H. 2009. Comparative genomics-guided loop-mediated isothermal amplification for characterization of Pseudomonas syringae pv. Phaseolicola. J. Appl. Microbiol. 107: 717-726. 32.Lister, R. M., Bancroft, J. B., and Nadakavukaren, M. J. 1965. Some sap-transmissible viruses from apple. Phytopathology 55: 859-870. 33.Lu, Y., Yao, B., Wang, G., and Hong, N. 2018. The detection of ACLSV and ASPV in pear plants by RT-LAMP assays. J. Virol. Methods 252: 80-85. 34.Martelli, G. P., Adams, M. J., Kreuze, J. F., and Dolja, V. V. 2007. Family Flexiviridae: a case study in virion and genome plasticity. Annu. Rev. Phytopathol. 45: 73-100. 35.Menzel, W., Jelkmann, W., and Maiss, E. 2002. Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control. J. Virol. Methods 99: 81-92. 36.Mori, Y., and Notomi, T. 2009. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J. Infect. Chemother. 15: 62-69. 37.Nie, X. 2005. Reverse transcription loop-mediated isothermal amplification of DNA for detection of Potato virus Y. Plant Disease 89 (6): 605-610. 38.Niu, J. H., Yang, D., Liu, Q., Guo, Y. D., Guo, Q. X., Jian, H., and Chen, C. L. 2011. Rapid detection of Meloidogyne spp. by LAMP assay in soil and roots. Crop Prot. 30 (8): 1063-1069. 39.Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., and Hase, T. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28 (12): E63. 40.Peng, J., Fan, Z. F., and Huang, J. S. 2012a. Rapid detection of Banana streak virus by loop-mediated isothermal amplification assay in South China. J. Phytopathol. 160: 248-250. 41.Peng, J., Shi, M. J., Xia, Z. H., Huang, J. S., and Fan, Z. F. 2012b. Detection of Cucumber mosaic virus isolates from banana by one step. Arch. Virol. 157: 2213-2217. 42.Peng, D., Xie, J., Qiang, W., Ling, K.S., Guo, L., Fan, Z., and Zhou, T. 2017. One-step reverse transcription loop-mediated isothermal amplification assay for detection of Apple chlorotic leaf spot virus. 248: 154-158. 43.Shiel, P. J., and Berger, P. H. 2000. The complete nucleotide sequence of Apple mosaic virus (ApMV) RNA 1 and RNA 2: ApMV is more closely related to Alfalfa mosaic virus than to other ilarviruses. J. Gen. Virol. 81: 273–278. 44.Shim, H. K., Hwang, K. H., Shim, C. K., Son, S. W., Kim, D., Choi, Y. M., Chung, Y., Kim, D. H., Jee, H. J., and Lee, S. C. 2006. The pear black necrotic leaf spot disease virus transmitted by Talaromyces flavus displays pathogenicity similar to Apple stem grooving virus strains. Plant Pathol. J. 22: 255-259. 45.TomLinson, J. A., Boonham, N., and Barker, I. 2007. Faster, simpler, more-specific methods for improved molecular detection of Phytophthora ramorum in the field. Appl. Environ. Microbiol. 73 (12): 4040-4047. 46.TomLinson, J. A., Boonham, N., and Dickinson, M. 2010a. Rapid detection of Phytophthora ramorum and P. kernoviae by two-minute DNA extraction followed by isothermal amplification and amplicon detection by generic lateral flow device. Phytopathology 100 (2): 143-149. 47.TomLinson, J. A., Boonham, N., and Dickinson, M. 2010b. Development and evaluation of a one-hour DNA extractionand loop-mediated isothermal amplification assay for rapid detection of phytoplasmas. Plant Pathol. 59: 465-471. 48.Watpade, S., Raigond, B., Handa, A., Bhardwaj, P., Pramanick, K.K., Verma, A., Kapoor, S., and Sharma, U. 2018. RT-LAMP detection of Apple chlorotic leaf spot virus in apple and pear. Plant Dis. Res. 33 (1): 94-98. 49.Wu, Z. B., Ku, H. M., Chen, Y. K., Chang, C. J., and Jan, F. J. 2010a. Biological and molecular characterization of Apple chlorotic leaf spot virus causing chlorotic leaf spot on pear (Pyrus pyrifolia) in Taiwan. HortScience 45: 1073–1078. 50.Wu, Z. B., Ku, H. M., Su, C. C., Chen, I. Z., and Jan, F. J. 2010b. Molecular and biological characterization of an isolate of Apple stem pitting virus causing pear vein yellow disease in Taiwan. J. Plant Pathol. 92: 721-728. 51.Wu, Z. B., Zheng, Y. X., Su, C. C., Chang, C. J., and Jan, F. J. 2010c. Identification and characterization of Apple stem grooving virus causing leaf distortion on pear (Pyrus pyrifolia) in Taiwan. Eur. J. Plant Pathol. 128: 71-79. 52.Zhang, Z. Y., Liu, X. J., Li, D. W., Yu, J. L., and Han, C. G. 2011. Rapid detection of Wheat yellow mosaic virus by reverse transcription loop-mediated isothermal amplification. Virol. J. 8: 550. 53.Zhao, L., Cheng, J. L., Hao, X. G., Tian, X. M., and Wu, Y. F. 2012. Rapid detection of tobacco viruses by reverse transcription loop-mediated isothermal amplification. Arch. Virol. 159 (12): 2291-2298. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21846 | - |
dc.description.abstract | 為生產高品質溫帶梨,臺灣每年冬季自日本及中國大陸進口帶有花芽之溫帶梨枝條作為接穗,嫁接於中低海拔橫山梨之徒長枝。蘋果黃化葉斑病毒 (Apple chlorotic leaf spot virus, ACLSV) 為國外普遍感染薔薇科果樹且造成經濟危害之重要植物病毒,因此被列為梨接穗輸入時須檢測之病原種類,現行 ACLSV 之檢測主要以酵素連結免疫吸附分析 (enzyme-linked immunosorbent assay, ELISA) 及反轉錄聚合酶連鎖反應 (reverse transcription polymerase chain reaction, RT-PCR) 為主,然此等方法需要特別儀器,且獲得偵測結果所需之時間仍長,有時亦會有偽陽性之問題產生。反轉錄恆溫環狀擴增法 (reverse transcription loop-mediated isothermal amplification, RT-LAMP) 是一種可在恆溫下反應且即時、簡便具高專一性及高效率之核酸增幅方法,可直接以肉眼辨識反應結果。為改善現行檢測效率,擬研發以 RT-LAMP 來檢測 ACLSV,首先合成 ACLSV不同病毒分離株之鞘蛋白基因 (coat protein gene, CP) DNA 為標的序列,並利用胞外轉錄合成 RNA 做為本試驗之正對照組病毒核酸,同時針對各鞘蛋白基因區域設計共4條之專一性引子組,並於恆溫63˚C下進行60分鐘增幅反應可順利偵測到 ACLSV,但對其他亦普遍感染梨樹之病毒如蘋果莖痘斑病毒 (Apple stem pitting virus) 及蘋果莖凹陷病毒 (Apple stem grooving virus) 則無法被增幅。為比較 RT-LAMP 與 RT-PCR 兩者之靈敏度,將 ACLSV 之 A4 分離株 RNA 經過10倍序列稀釋並混合健康梨 RNA 後,同時進行此兩反應,發現 RT-LAMP 於增幅70分鐘時其靈敏度較 RT-PCR 高出100倍。而比較 RT-LAMP 引子組對三分離株增幅之靈敏度差異,發現 A4 分離株增幅之靈敏度較 Kuerle 及 YT-2-CS 分離株高,可歸因於引子組對於 A4 分離株有較佳之黏合性。接著實際以所建立之 RT-LAMP 技術對2013至2014年輸入的其中473枝梨接穗樣本進行篩檢,並輔以 ELISA 及 RT-PCR 技術做重複確認,發現得到一致結果。綜合以上研究,RT-LAMP 較現行方法縮短檢測時程,提高檢測效率,兼具專一及靈敏的特性,期能藉以對輸入梨接穗之 ACLSV 提供更快速、方便、準確且靈敏之檢測方式。 | zh_TW |
dc.description.abstract | To produce high quality pear, branches with flower buds of pear grown in temperate region are imported from Japan and China to serves as scions to suckers of Hengshan pear in Taiwan. However, an important plant virus, Apple chlorotic leaf spot virus (ACLSV), commonly infects Rosaceae fruit trees and causes economic damages in foreign countries. Therefore, ACLSV has been listed as a quarantine pathogen. Currently, the detection of ACLSV is mainly conducted by use of enzyme-linked immunesorbent assay (ELISA) or reverse transcription polymerase chain reaction (RT-PCR). However, both ELISA and RT-PCR require specific equipments and are also time-consuming. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a real-time, simple and highly efficient method to amplify nucleic acid and can be processed under an isothermal condition. Furthermore, the result of RT-LAMP can be observed by change of colors without a specific equipment. To improve current detection methods, this research plans to develop RT-LAMP for the detection of ACLSV. We synthesized DNA of the coat protein (CP) genes derived from three isolates of ACLSV, and use in vitro transcription to generate ACLSV RNA as the positive control. We have optimized the RT-LAMP detection, and the amplification of ACLSV can be achieved at 63˚C for 60 minutes using a set of 4 specific primers targeting the viral CP genes. In addition, the RT-LAMP can only detect ACLSV but not viruses commonly infect pear such as Apple stem pitting virus and Apple stem grooving virus. To compare the sensitivity of RT-LAMP with that of RT-PCR, RNA isolated from A4 isolates of ACLSV was diluted serially in 10-fold increments and added to RNA from healthy pear for the assays. After 70 minutes of RT-LAMP reaction, the result showed that the detection sensitivity of the RT-LAMP was 100 times higher than that of the RT-PCR. Besides, comparing the detection sensitivity of the RT-LAMP primers to the three isolates of ACLSV, the result showed that A4 isolate was higher than that of Kuerle and YT-2-CS isolates, which was attributed to better combination ability of the primers to the sequence of A4 isolate. A total of 473 pear scions imported from 2013 to 2014 were screened by the developed RT-LAMP and confirmed by ELISA and RT-PCR. The results of ACLSV detection by use of the three methods were consistent. It suggests that the developed RT-LAMP is convenient and time-saving without sacrificing specificity and sensitivity. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T03:49:29Z (GMT). No. of bitstreams: 1 ntu-107-R00645012-1.pdf: 5544958 bytes, checksum: 57b12ad6cff7ebb05776d97e95393c60 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 誌謝 I
中文摘要 II Abstract IV 目錄 VI 表目錄 IX 圖目錄 X 壹、前言 1 貳、前人文獻探討 3 一、梨樹栽培概況 3 二、臺灣梨接穗檢疫法規及輸入概況 4 (一) 臺灣梨接穗檢疫法規沿革 4 (二) 臺灣梨接穗近年輸入概況 6 三、感染梨樹重要病毒種類及其特性 6 (一) 蘋果黃化葉斑病毒 (Apple chlorotic leaf spot virus, ACLSV) 6 (二) 蘋果莖痘斑病毒 (Apple stem pitting virus, ASPV) 8 (三) 蘋果莖凹陷病毒 (Apple stem grooving virus, ASGV) 9 (四) 蘋果嵌紋病毒 (Apple mosaic virus, ApMV) 10 (五) 梨潛隱病毒 (Peach latent mosaic viroid, PLMVd) 10 (六) 番茄叢生矮化病毒 (Tomato bushy stunt virus, TBSV) 10 四、蘋果黃化葉斑病毒 (ACLSV) 之風險潛能 (Pest risk potential) 11 (一) 傳入之後果評估 11 (二) 傳入可能性之風險評等 12 (三) 蘋果黃化葉斑病毒 (ACLSV) 之風險潛能評估結果 13 五、現行輸入梨接穗常用病毒檢測技術 14 (一) 血清學檢測技術 14 (二) 核酸增幅技術 14 (三) 現行檢測技術待改善之處及研究動機 14 六、恆溫環狀擴增法之原理及特性 15 七、應用恆溫環狀擴增法檢測植物病原之實例 16 (一) 病毒與類病毒 16 (二) 真菌 17 (三) 細菌 18 (四) 植物菌質體 18 (五) 線蟲 19 參、材料與方法 20 一、供試植物檢測材料 20 二、供試目標病毒製備 20 (一) 篩選欲合成之 ACLSV 分離株 20 (二) 合成病毒鞘蛋白核酸序列 21 (三) 擴增所合成病毒鞘蛋白核酸序列及保存 21 (四) 抽取質體 DNA 及確認目標基因 21 (五) 限制酶作用及核酸純化 22 (六) 生體外轉錄 (In vitro Transcription) 及 RNA 轉錄本之純化與保存 23 三、RT-LAMP 專一性引子組之設計 24 四、引子組檢測之反應條件及環境測試 24 五、混合病毒與植物核酸同時進行 ACLSV 及梨之 actin gene檢測 25 六、評估 RT-LAMP 檢測 ACLSV 病毒之專一性 25 七、評估 RT-LAMP 偵測 ACLSV 病毒之靈敏度 26 八、對輸入梨接穗樣本進行病毒檢測 26 (一) 植物全 RNA 之純化 26 (二) 反轉錄恆溫環狀擴增法 (RT-LAMP) 27 (三) 反轉錄聚合酶連鎖反應 (RT-PCR) 28 (四) ELISA 檢測 29 肆、結果 31 一、目標病毒序列分析以篩選欲合成之 ACLSV 分離株 31 二、抽取質體 DNA 後確認轉型成功 31 三、以限制酶作用確認 ACLSV 之鞘蛋白基因選殖成功 32 四、供試病毒之 RNA 轉錄本 (transcript) 製備 32 五、檢測 ACLSV 之 RT-LAMP 引子組設計 32 六、檢測 ACLSV 引子組之反應條件及環境測試 32 七、檢測梨肌動蛋白基因 (actin gene) 之 RT-LAMP 引子組………………..33 八、混合病毒與植物核酸同時進行 ACLSV 及梨之 actin gene 檢測………33 九、評估 RT-LAMP 檢測 ACLSV 之專一性 34 十、評估 RT-LAMP 檢測 ACLSV 之靈敏度 34 十一、以RT-PCR 檢測 ACLSV 35 十二、以 RT-PCR 檢測梨肌動蛋白基因 (actin gene) 35 十三、輸入梨接穗檢疫時採樣之樣本進行病毒檢測 35 伍、討論 38 陸、結論 81 柒、參考文獻 82 | |
dc.language.iso | zh-TW | |
dc.title | 應用反轉錄恆溫環狀擴增法偵測輸入梨接穗之蘋果黃化葉斑病毒 | zh_TW |
dc.title | The detection of Apple chlorotic leaf spot virus from imported pear scions by the method of reverse transcription loop-mediated isothermal amplification (RT-LAMP) | en |
dc.type | Thesis | |
dc.date.schoolyear | 107-1 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 洪挺軒(Ting-Hsuan Hung) | |
dc.contributor.oralexamcommittee | 吳文哲(Wen-Jer Wu),林乃君(Nai-Chun Lin),鍾嘉綾(Chia-Lin Chung) | |
dc.subject.keyword | 蘋果黃化葉斑病毒,梨接穗,反轉錄恆溫環狀擴增法,鞘蛋白基因,酵素連結免疫吸附分析,反轉錄聚合?連鎖反應, | zh_TW |
dc.subject.keyword | Apple chlorotic leaf spot virus (ACLSV),pear scion,reverse transcription loop-mediated isothermal amplification (RT-LAMP),coat protein gene,enzyme-linked immunosorbent assay (ELISA),reverse transcription polymerase chain reaction (RT-PCR), | en |
dc.relation.page | 88 | |
dc.identifier.doi | 10.6342/NTU201804321 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2018-12-06 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 植物醫學碩士學位學程 | zh_TW |
顯示於系所單位: | 植物醫學碩士學位學程 |
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ntu-107-1.pdf 目前未授權公開取用 | 5.41 MB | Adobe PDF |
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