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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王錦堂 | |
| dc.contributor.author | Po-An Su | en |
| dc.contributor.author | 蘇柏安 | zh_TW |
| dc.date.accessioned | 2021-06-16T08:04:03Z | - |
| dc.date.available | 2019-10-09 | |
| dc.date.copyright | 2014-10-09 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-07-02 | |
| dc.identifier.citation | 1. Ryan, K. R., CG, eds. (2004) Sherris Medical Microbiology(4th ed.). McGraw Hill. ISBN 0-8385-8529-9.
2. Podschun, R., and Ullmann, U. (1998) Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603 3. Liu, Y. C., Cheng, D. L., and Lin, C. L. (1986) Klebsiella pneumoniae liver abscess associated with septic endophthalmitis. Arch Intern Med 146, 1913-1916 4. Saccente, M. (1999) Klebsiella pneumoniae liver abscess, endophthalmitis, and meningitis in a man with newly recognized diabetes mellitus. Clin Infect Dis 29, 1570-1571 5. Wang, J. H., Liu, Y. C., Lee, S. S., Yen, M. Y., Chen, Y. S., Wann, S. R., and Lin, H. H. (1998) Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438 6. Yang, C. C., Yen, C. H., Ho, M. W., and Wang, J. H. (2004) Comparison of pyogenic liver abscess caused by non-Klebsiella pneumoniae and Klebsiella pneumoniae. J Microbiol Immunol Infect 37, 176-184 7. Tsai, F. C., Huang, Y. T., Chang, L. Y., and Wang, J. T. (2008) Pyogenic liver abscess as endemic disease, Taiwan. Emerg Infect Dis 14, 1592-1600 8. Ko, W. C., Paterson, D. L., Sagnimeni, A. J., Hansen, D. S., Von Gottberg, A., Mohapatra, S., Casellas, J. M., Goossens, H., Mulazimoglu, L., Trenholme, G., Klugman, K. P., McCormack, J. G., and Yu, V. L. (2002) Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. Emerg Infect Dis 8, 160-166 9. Yeoh, K. G., Yap, I., Wong, S. T., Wee, A., Guan, R., and Kang, J. Y. (1997) Tropical liver abscess. Postgrad Med J 73, 89-92 10. Garcia-Morillo, J. S., de la Cruz-Vicente, F., Lopez-Ruiz, T., and Bernabeu-Wittel, M. (2006) Hepatic abscess due to Klebsiella pneumoniae in an Asian immigrant. Enferm Infecc Microbiol Clin 24, 585-586 11. Batres Iglesias, A. P., Perez Cabeza, M. I., Del Rio Pardo, M. J., and Castano, M. (2011) Endogenous endophthalmitis as a first clinical manifestation of Klebsiella sepsis. The importance of an early diagnosis. Arch Soc Esp Oftalmol 86, 412-414 12. Gomez, C. B., Amparo; Otero, Joaquin R.; Chaves, Fernando (2007) Primary pyogenic liver abscess caused by magA? Klebsiella pneumoniae in Spain. Clinical Microbiology Newsletter 29, 100-103 13. Karama, E. M., Willermain, F., Janssens, X., Claus, M., Van den Wijngaert, S., Wang, J. T., Verougstraete, C., and Caspers, L. (2008) Endogenous endophthalmitis complicating Klebsiella pneumoniae liver abscess in Europe: case report. Int Ophthalmol 28, 111-113 14. Meric de Bellefon, L., Legrand, J. C., Codden, T., Carlier, E., and Vanhaeverbeek, M. (2007) Klebsiella pneumoniae septicaemia and meningitis in a diabetic patient with an hepatic abscess. Rev Med Brux 28, 460-463 15. Moffie, B. G., and Wijbenga, J. A. (2008) Liver abscess due to Klebsiella pneumoniae showing the mucoid phenotype. J Infect 57, 353-354 16. Sobirk, S. K., Struve, C., and Jacobsson, S. G. (2010) Primary Klebsiella pneumoniae Liver Abscess with Metastatic Spread to Lung and Eye, a North-European Case Report of an Emerging Syndrome. Open Microbiol J 4, 5-7 17. Casella, F., Finazzi, L., Repetti, V., Rubin, G., Dimarco, M., Mauro, T., and Furlan, R. (2009) Liver abscess caused by Klebsiella pneumoniae: two case reports. Cases J 2, 6879 18. Merlet A, C. C., Dutronc H, de Barbeyrac B, Brisse S, Dupon M. (2012) Primary liver abscess due to CC23-K1 virulent clone of Klebsiella pneumoniae in France. Clin Microbiol Infect 18, 338-339 19. Decre, D., Verdet, C., Emirian, A., Le Gourrierec, T., Petit, J. C., Offenstadt, G., Maury, E., Brisse, S., and Arlet, G. (2011) Emerging severe and fatal infections due to Klebsiella pneumoniae in two university hospitals in France. J Clin Microbiol 49, 3012-3014 20. Anstey, J. R., Fazio, T. N., Gordon, D. L., Hogg, G., Jenney, A. W., Maiwald, M., and Wilksch, J. J. (2010) Community-acquired Klebsiella pneumoniae liver abscesses - an 'emerging disease' in Australia. Med J Aust 193, 543-545 21. Kauffmann, F. (1949) On The Serology Of The Klebsiella Group. Acta Pathologica Microbiologica Scandinavica 26, 381-406 22. Pan, Y. J., Fang, H. C., Yang, H. C., Lin, T. L., Hsieh, P. F., Tsai, F. C., Keynan, Y., and Wang, J. T. (2008) Capsular polysaccharide synthesis regions in Klebsiella pneumoniae serotype K57 and a new capsular serotype. J Clin Microbiol 46, 2231-2240 23. Hsu, C. R., Lin, T. L., Pan, Y. J., Hsieh, P. F., and Wang, J. T. (2013) Isolation of a bacteriophage specific for a new capsular type of Klebsiella pneumoniae and characterization of its polysaccharide depolymerase. PLoS One 8, e70092 24. Fung, C. P., Chang, F. Y., Lee, S. C., Hu, B. S., Kuo, B. I., Liu, C. Y., Ho, M., and Siu, L. K. (2002) A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut 50, 420-424 25. Amako, K., Meno, Y., and Takade, A. (1988) Fine structures of the capsules of Klebsiella pneumoniae and Escherichia coli K1. J Bacteriol 170, 4960-4962 26. Alvarez, D., Merino, S., Tomas, J. M., Benedi, V. J., and Alberti, S. (2000) Capsular polysaccharide is a major complement resistance factor in lipopolysaccharide O side chain-deficient Klebsiella pneumoniae clinical isolates. Infect Immun 68, 953-955 27. Clements, A., Gaboriaud, F., Duval, J. F., Farn, J. L., Jenney, A. W., Lithgow, T., Wijburg, O. L., Hartland, E. L., and Strugnell, R. A. (2008) The major surface-associated saccharides of Klebsiella pneumoniae contribute to host cell association. PLoS One 3, e3817 28. Lin, J. C., Chang, F. Y., Fung, C. P., Xu, J. Z., Cheng, H. P., Wang, J. J., Huang, L. Y., and Siu, L. K. (2004) High prevalence of phagocytic-resistant capsular serotypes of Klebsiella pneumoniae in liver abscess. Microbes Infect 6, 1191-1198 29. Bradford, P. A. (2001) Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin Microbiol Rev 14, 933-951 30. Chaves, J., Ladona, M. G., Segura, C., Coira, A., Reig, R., and Ampurdanes, C. (2001) SHV-1 beta-lactamase is mainly a chromosomally encoded species-specific enzyme in Klebsiella pneumoniae. Antimicrob Agents Chemother 45, 2856-2861 31. Jacoby, G. A., and Medeiros, A. A. (1991) More extended-spectrum beta-lactamases. Antimicrob Agents Chemother 35, 1697-1704 32. Yu, W. L., Chuang, Y. C., and Walther-Rasmussen, J. (2006) Extended-spectrum beta-lactamases in Taiwan: epidemiology, detection, treatment and infection control. J Microbiol Immunol Infect 39, 264-277 33. Yan, J. J., Hsueh, P. R., Lu, J. J., Chang, F. Y., Shyr, J. M., Wan, J. H., Liu, Y. C., Chuang, Y. C., Yang, Y. C., Tsao, S. M., Wu, H. H., Wang, L. S., Lin, T. P., Wu, H. M., Chen, H. M., and Wu, J. J. (2006) Extended-spectrum beta-lactamases and plasmid-mediated AmpC enzymes among clinical isolates of Escherichia coli and Klebsiella pneumoniae from seven medical centers in Taiwan. Antimicrob Agents Chemother 50, 1861-1864 34. Mendes, R. E., Mendoza, M., Banga Singh, K. K., Castanheira, M., Bell, J. M., Turnidge, J. D., Lin, S. S., and Jones, R. N. (2013) Regional Resistance Surveillance Program Results for Twelve Asia-Pacific Nations (2011). Antimicrob Agents Chemother 35. Woodford, N., Dallow, J. W., Hill, R. L., Palepou, M. F., Pike, R., Ward, M. E., Warner, M., and Livermore, D. M. (2007) Ertapenem resistance among Klebsiella and Enterobacter submitted in the UK to a reference laboratory. Int J Antimicrob Agents 29, 456-459 36. Doumith, M., Ellington, M. J., Livermore, D. M., and Woodford, N. (2009) Molecular mechanisms disrupting porin expression in ertapenem-resistant Klebsiella and Enterobacter spp. clinical isolates from the UK. J Antimicrob Chemother 63, 659-667 37. Chiu, S. K., Wu, T. L., Chuang, Y. C., Lin, J. C., Fung, C. P., Lu, P. L., Wang, J. T., Wang, L. S., Siu, L. K., and Yeh, K. M. (2013) National Surveillance Study on Carbapenem Non-Susceptible Klebsiella pneumoniae in Taiwan: The Emergence and Rapid Dissemination of KPC-2 Carbapenemase. PLoS One 8, e69428 38. Nordmann, P., Cuzon, G., and Naas, T. (2009) The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria. Lancet Infect Dis 9, 228-236 39. (2011) Annual report of Taiwan Nosocomial Infection Surveillance Systems. http://www.cdc.gov.tw/english/index.aspx 40. Castanheira, M., Sader, H. S., Deshpande, L. M., Fritsche, T. R., and Jones, R. N. (2008) Antimicrobial activities of tigecycline and other broad-spectrum antimicrobials tested against serine carbapenemase- and metallo-beta-lactamase-producing Enterobacteriaceae: report from the SENTRY Antimicrobial Surveillance Program. Antimicrob Agents Chemother 52, 570-573 41. Tsai, S. S., Huang, J. C., Chen, S. T., Sun, J. H., Wang, C. C., Lin, S. F., Hsu, B. R., Lin, J. D., Huang, S. Y., and Huang, Y. Y. (2010) Characteristics of Klebsiella pneumoniae bacteremia in community-acquired and nosocomial infections in diabetic patients. Chang Gung Med J 33, 532-539 42. d'Herelle, F. (1931) Bacteriophage as a Treatment in Acute Medical and Surgical Infections. Bull N Y Acad Med 7, 329-348 43. Slopek, S., Durlakowa, I., Weber-Dabrowska, B., Kucharewicz-Krukowska, A., Dabrowski, M., and Bisikiewicz, R. (1983) Results of bacteriophage treatment of suppurative bacterial infections. I. General evaluation of the results. Arch Immunol Ther Exp (Warsz) 31, 267-291 44. Slopek, S., Durlakowa, I., Weber-Dabrowska, B., Kucharewicz-Krukowska, A., Dabrowski, M., and Bisikiewicz, R. (1983) Results of bacteriophage treatment of suppurative bacterial infections. II. Detailed evaluation of the results. Arch Immunol Ther Exp (Warsz) 31, 293-327 45. Slopek, S., Kucharewicz-Krukowska, A., Weber-Dabrowska, B., and Dabrowski, M. (1985) Results of bacteriophage treatment of suppurative bacterial infections. VI. Analysis of treatment of suppurative staphylococcal infections. Arch Immunol Ther Exp (Warsz) 33, 261-273 46. Slopek, S., Kucharewicz-Krukowska, A., Weber-Dabrowska, B., and Dabrowski, M. (1985) Results of bacteriophage treatment of suppurative bacterial infections. V. Evaluation of the results obtained in children. Arch Immunol Ther Exp (Warsz) 33, 241-259 47. Slopek, S., Kucharewicz-Krukowska, A., Weber-Dabrowska, B., and Dabrowski, M. (1985) Results of bacteriophage treatment of suppurative bacterial infections. IV. Evaluation of the results obtained in 370 cases. Arch Immunol Ther Exp (Warsz) 33, 219-240 48. Slopek, S., Weber-Dabrowska, B., Dabrowski, M., and Kucharewicz-Krukowska, A. (1987) Results of bacteriophage treatment of suppurative bacterial infections in the years 1981-1986. Arch Immunol Ther Exp (Warsz) 35, 569-583 49. Weber-Dabrowska, B., Mulczyk, M., and Gorski, A. (2003) Bacteriophages as an efficient therapy for antibiotic-resistant septicemia in man. Transplant Proc 35, 1385-1386 50. Weber-Dabrowska, B., Mulczyk, M., and Gorski, A. (2000) Bacteriophage therapy of bacterial infections: an update of our institute's experience. Arch Immunol Ther Exp (Warsz) 48, 547-551 51. Wittebole, X., De Roock, S., and Opal, S. M. (2013) A historical overview of bacteriophage therapy as an alternative to antibiotics for the treatment of bacterial pathogens. Virulence 4 52. Bayer, M. E., Thurow, H., and Bayer, M. H. (1979) Penetration of the polysaccharide capsule of Escherichia coli (Bi161/42) by bacteriophage K29. Virology 94, 95-118 53. Hildegard Geyer, K. H., Bartłomiej Kwiatkowski, S. Schlecht and S. Stirm. (1983) Degradation of bacterial surface carbohydrates by virus-associated enzymes. Pure and Applied Chemistry 55, 637-653 54. Gaston, M. A., Ayling-Smith, B. A., and Pitt, T. L. (1987) New bacteriophage typing scheme for subdivision of the frequent capsular serotypes of Klebsiella spp. J Clin Microbiol 25, 1228-1232 55. Pieroni, P. (1996) Thesis: Application of Bacteriophage Typing to Klebsiella pneumonia. Saskatchewan University, Saskatchewan, Canada. 56. Mushtaq, N., Redpath, M. B., Luzio, J. P., and Taylor, P. W. (2005) Treatment of experimental Escherichia coli infection with recombinant bacteriophage-derived capsule depolymerase. J Antimicrob Chemother 56, 160-165 57. Chang, H. C., Chen, C. R., Lin, J. W., Shen, G. H., Chang, K. M., Tseng, Y. H., and Weng, S. F. (2005) Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. App Environ Microbiol 71, 1387-1393 58. Briandet, R., Lacroix-Gueu, P., Renault, M., Lecart, S., Meylheuc, T., Bidnenko, E., Steenkeste, K., Bellon-Fontaine, M. N., and Fontaine-Aupart, M. P. (2008) Fluorescence correlation spectroscopy to study diffusion and reaction of bacteriophages inside biofilms. App Environ Microbiol 74, 2135-2143 59. Chibani-Chennoufi, S., Dillmann, M. L., Marvin-Guy, L., Rami-Shojaei, S., and Brussow, H. (2004) Lactobacillus plantarum bacteriophage LP65: a new member of the SPO1-like genus of the family Myoviridae. J Bacteriol 186, 7069-7083 60. Wu, L. T., Chang, S. Y., Yen, M. R., Yang, T. C., and Tseng, Y. H. (2007) Characterization of extended-host-range pseudo-T-even bacteriophage Kpp95 isolated on Klebsiella pneumoniae. App Environ Microbiol 73, 2532-2540 61. 陳青青. (2013.7) 以噬菌體重組基因技術探討多宿主噬菌體K11-7-1-1之莢膜分解酶酵素活性. 國立台灣大學醫學院微生物學研究所 碩士論文 62. Chuang, Y. P., Fang, C. T., Lai, S. Y., Chang, S. C., and Wang, J. T. (2006) Genetic determinants of capsular serotype K1 of Klebsiella pneumoniae causing primary pyogenic liver abscess. J Infect Dis 193, 645-654 63. Huyan, X. H., Lin, Y. P., Gao, T., Chen, R. Y., and Fan, Y. M. (2011) Immunosuppressive effect of cyclophosphamide on white blood cells and lymphocyte subpopulations from peripheral blood of Balb/c mice. International immunopharmacology 11, 1293-1297 64. Lin, T. L., Huang, Y.T, Lee, W.C., Tsai, Y.T., Su, P.A., Pan, Y.J., Hsu, C.R., Wu, M.C., Wang, J.T. (2014) Isolation of a bacteriophage and its depolymerase specific for K1 capsule of Klebsiella pneumoniae: implication in typing and treatment. J Infect Dis in press 65. Pennington, J. E., Matthews, W. J., Jr., Marino, J. T., Jr., and Colten, H. R. (1979) Cyclophosphamide and cortisone acetate inhibit complement biosynthesis by guinea pig bronchoalveolar macrophages. J. Immunol 123, 1318-1321 66. Franklin, G. A., Scott, M. J., Patel, M., Hoth, J. J., Peyton, J. C., and Cheadle, W. G. (2003) A novel model of pneumonia from intraperitoneal injection of bacteria. American journal of surgery 186, 493-499 67. Kumari, S., Harjai, K., and Chhibber, S. (2011) Bacteriophage versus antimicrobial agents for the treatment of murine burn wound infection caused by Klebsiella pneumoniae B5055. J Med Microbiol 60, 205-210 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57972 | - |
| dc.description.abstract | 近年來，克雷伯氏肺炎桿菌造成的社區侵襲性原發性肝膿瘍成為了全球性的新興感染症。克雷伯氏肺炎桿菌菌體外的莢膜是重要的致病因子之一，可以抵抗宿主免疫細胞吞噬作用以及補體毒殺機制，而在台灣造成原發性肝膿瘍以K1、K2莢膜血清型最為常見，充分顯示莢膜型和致病力的相關性。克雷伯氏肺炎桿菌菌株的多重抗藥性問題日趨嚴重，例如Extended-spectrumβ-lactamase(ESBL)基因及Carbapenem-resistant Klebsiella pneumoniae(CRKP)。先前本實驗室已經找到能夠專一性感染K1 莢膜型菌株的噬菌體NTUH-K2044-K1-1 並具有全基因體定序，從中找到一為K1 莢膜分解酶的基因產物。本研究欲評估莢膜分解酶治療效果，因此接著篩選出出感染K2 莢膜型菌株的噬菌體1611E-K2-1 並從中找出K2 莢膜分解酶基因，利用基因重組技術大量表現純化出K1、K2 莢膜分解酶並證實對小鼠不具毒性，以腹腔注射感染K1、K2 莢膜型菌株的小鼠在感染後1 小時內接受單劑莢膜分解酶（25 μg）治療可提高87.5%（K1）或100%（K2）的存活率。目前將莢膜分解酶療法擴展至多重抗藥性菌株感染的治療，實驗室統計台大、長庚、成大和榮總四家醫院的CRKP 臨床菌株發現有集中於K64 莢膜型的現象。欲模擬臨床上易感染多重抗藥性菌株之免疫不全病人族群，利用Cyclophosphamide 建立免疫不全小鼠模式，以腹腔注射感染K64 莢膜型菌株後1 小時接受單劑K64 莢膜分解酶（150 μg）進行治療，仍可成功提高62.5%的存活率。因此，基因表現之莢膜分解酶應可作為傳統抗生素之外治療克雷伯氏肺炎桿菌感染的替代療法。 | zh_TW |
| dc.description.abstract | Community-acquired pyogenic liver abscess (PLA) caused by Klebsiella pneumoniae has become an emerging infectious disease. The capsule of K. pneumoniae is an important virulence factor, which can keep bacteria from phagocytosis and complement mediated lysis. Previous studies reported K1 and K2 were prevalent capsular serotypes of strains causing PLA in Taiwan. Multi-drug resistances of K. pneumoniae including Extended-spectrumβ-lactam resistance and Carbapenem-resistance Klebsiella pneumoniae (CRKP) are urgent global issues. Phage NTUH-K2044-K1-1 and its capsule depolymerase which were both specific to K1 capsule were identified in our previous study. Here, we isolated a phage 1611E-K2-1 shown to infect K2 strains and identified its K2 capsule depolymerase. Intraperitoneal administration of a single dose of 25 ug K1 or K2 capsule depolymerase at 1h after intraperitoneal infection of K. pneumoniae K1 or K2 strain could enhance 87.5 %(K1) or 100 %(K2) survival rate, respectively. We further tested capsule depolymerase treatment of multi-drug resistant (MDR) K. pneumoniae infection. We observed the most prevalent capsular type of CRKP from NTUH, CGUH, NCKUH, and VGH was K64. The efficacy of K64 capsule depolymerase treatment in cyclophosphamide-treated mice mimicing the immune-compromised patients infected with MDR K. pneumoniae was evaluated. Intraperitoneal administration of a single dose of 150 ug K64 capsule depolymerase at 1h after intraperitoneal infection of K. pneumoniae K64 strain also improve 62.5 % survival. Therefore, capsule depolymerase treatment is a potential therapy for K. pneumoniae infection alternative to conventional antibiotics treatment. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T08:04:03Z (GMT). No. of bitstreams: 1 ntu-103-R01445126-1.pdf: 4878548 bytes, checksum: e90430782903810fb9f04ba3fdcfb284 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 目錄
口試委員審定書 I 致謝 II 中文摘要 III Abstract IV 第一章、緒論 1 1-1 全球性新興感染症：克雷伯氏肺炎桿菌造成之原發性肝膿瘍 1 1-2 克雷伯氏肺炎桿菌的莢膜血清分型 2 1-3 克雷伯氏肺炎桿菌的多重抗藥性菌株（multidrug resistant, MDR） 2 1-4 多重抗藥性克雷伯氏肺炎桿菌感染的抗生素治療 4 1-5 尋找新的替代療法（Alternative therapy）取代傳統抗生素治療 5 1-6 研究目的 9 第二章、材料與方法 10 2-1 實驗菌株 10 2-2 培養基 10 2-3 尋找及分離噬菌體 10 2-4 塗點試驗（Spot test） 11 2-5 噬菌體增生（Phage amplification） 11 2-6 噬菌體溶菌斑效價分析 12 2-7 噬菌體純化與DNA萃取 12 2-8 限制性核酸內切酶實驗 14 2-9 噬菌體全基因體序列分析 14 2-10 基因選殖（Cloning） 15 2-11 蛋白質純化與表現 16 2-12 SDS膠體電泳與考馬斯亮藍（coomassie brilliant blue）蛋白質定量法 18 2-13 莢膜多醣抽取法 20 2-14 莢膜多醣與莢膜分解酶作用實驗 21 2-15 實驗動物 22 2-16 以莢膜分解酶治療動物感染模式 22 2-17 西方墨點法（Western Blot） 22 2-18 血清毒殺試驗分析 23 2-19 建立免疫不全小鼠模式 24 2-20 以莢膜分解酶治療免疫不全小鼠感染模式 24 第三章、結果 25 3-1 尋找及分離可感染K2莢膜型克雷伯氏肺炎桿菌之K2噬菌體 25 3-2 1611E-K2-1溶菌斑效價分析及噬菌體增生 25 3-3 1611E-K2-1宿主範圍分析與對克雷伯氏肺炎桿菌K2莢膜型不同菌株之感染能力分析 26 3-4 獲取1611E-K2-1基因體 27 3-5 1611E-K2-1基因體定序與生物資訊分析 27 3-6 利用基因重組技術表現並純化K1-ORF33及K2-ORF16之基因產物 28 3-7 測試純化出的K1-ORF33及K2-ORF16基因產物酵素活性（enzyme activity） 29 3-8 確認K2莢膜分解酶對K2莢膜的降解切割能力 30 3-9 評估K1、K2莢膜分解酶治療對動物的安全性及副作用 30 3-10 評估K1、K2莢膜分解酶療法（capsule depolymerase treatment）的有效性 31 3-11 分析由臨床分離之抗藥性克雷伯氏肺炎桿菌菌株 31 3-12 測定克雷伯氏肺炎桿菌抗藥性菌株感染BALB/c小鼠的半致死劑量（LD50） 32 3-13 純化由噬菌體分離之K64莢膜分解酶 33 3-14 測試純化後之K64莢膜分解酶酵素活性 33 3-15 進行血清毒殺試驗分析（serum killing assay） 33 3-16 利用免疫抑制藥物（Cyclophosphamide）建立免疫不全小鼠模式並評估K64莢膜分解酶療法的有效性 34 第四章、討論 36 參考文獻 65 圖目錄 圖 一、確認K1莢膜分解酶蛋白質純化情形 42 圖 二、確認K2莢膜分解酶蛋白質純化情形 43 圖 三、K2莢膜分解酶對K2莢膜型菌株進行酵素效力試驗 44 圖 四、K2莢膜分解酶對K13莢膜型菌株進行酵素效力試驗 45 圖 五、確認K2莢膜分解酶對K2莢膜多醣的切割降解能力 46 圖 六、以西方墨點法確認注射過K1莢膜分解酶小鼠之血清抗體產生情形 47 圖 七、以西方墨點法確認注射過K2莢膜分解酶小鼠之血清抗體產生情形 48 圖 八、K1莢膜分解酶療法於NTUH-K2044（K1）克雷伯氏肺炎桿菌菌株感染BALB/c小鼠的療效評估 49 圖 九、K2莢膜分解酶療法於A4528（K2）克雷伯氏肺炎桿菌菌株感染BALB/c小鼠的療效評估 50 圖 十、分析K64莢膜型克雷伯氏肺炎桿菌ESBL臨床菌株的限制性片段長度多態性（restriction fragment length polymorphism, RFLP） 51 圖 十一、分析K64莢膜型克雷伯氏肺炎桿菌CRKP臨床菌株的限制性片段長度多態性（restriction fragment length polymorphism, RFLP） 52 圖 十二、確認K64莢膜分解酶蛋白質純化情形 53 圖 十三、K64莢膜分解酶對K64莢膜型菌株進行酵素效力試驗 54 圖 十四、血清毒殺試驗 55 圖 十五、以Cyclophosphamide藥物建立免疫不全小鼠模式期間小鼠體重變化 56 圖 十六、K64莢膜分解酶療法於7960（K64）克雷伯氏肺炎桿菌菌株感染免疫不全BALB/c小鼠的療效評估 57 表目錄 表 一、本研究使用細菌菌株及質體 58 表 二、研究中使用之引子（primer）DNA序列 59 表 三、K2噬菌體（1611E-K2-1）宿主範圍 60 表 四、National center for biotechnology information Basic Local Alignment Search Tool(NCBI BLAST)胺基酸序列資料庫，比對K2噬菌(1611E-K2-1)genome上ORFs序列之結果 61 表 五、噬菌體NTUH-K2044-K1-1之開放閱讀框K1-ORF33（K1莢膜分解酶基因序列）（1953 bp） 62 表 六、噬菌體1611E-K2-1之開放閱讀框K2-ORF16（K2莢膜分解酶基因序列）（2004 bp） 63 表 七、噬菌體K11-7-1-1之開放閱讀框S2-5（K64莢膜分解酶基因序列）（2988 bp） 64 | |
| dc.language.iso | zh-TW | |
| dc.title | 莢膜分解酶治療克雷伯氏肺炎桿菌感染之小鼠模式 | zh_TW |
| dc.title | Utilization of capsule depolymerase as a treatment for Klebsiella Pneumoniae infection in mice | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林稚容,董馨蓮,楊宏志,蔡丰喬 | |
| dc.subject.keyword | 克雷伯氏肺炎桿菌,莢膜型,噬菌體,多重抗藥性,莢膜分解?療法, | zh_TW |
| dc.subject.keyword | Klebsiella pneumoniae,capsule type,phage,multi-drug resistant,capsule depolymerase treatment, | en |
| dc.relation.page | 71 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-07-02 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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