臺灣博碩士論文加值系統

本研究是使用多套式PCR方法，以1個通用的反向引子和設計的8個具有種特異性的正向引子，快速鑑定臨床上陽性血液培養最常見的8種酵母菌。這8個種特異性引子設計的位置是位於核醣體DNA內轉錄區（internal transcribed spacer region, ITS）1和2，而通用性的引子則位於26S核醣體基因。這8種酵母菌經由多套式PCR方法所放大出來的DNA產物長度分別為：Candida lusitaniae (116 bp), C. parapsilosis (126 bp), C. tropicalis (149 bp), C. guilliermondii (185 bp), C. albicans (402~403 bp), C. krusei (475 bp), Cryptococcus neofonmans (516 bp), 和C. glabrata (631~632 bp)，而這些PCR產物經由膠體電泳即可有效將它們分離。以本法測試上述8種酵母菌的臨床分離共248株純菌，所獲得的敏感度（sensitivity）為100%（248/248），另外測試其他酵母菌及黴菌純菌（含36屬、71種、168株），所獲得的特異性（specificity）為98.2%（165/168）。再以此方法實際測試234個陽性血瓶（237分離株），這些菌種（菌株數）之分佈如下：C. albicans (121), C. tropicalis (50), C. glabrata (29), C. parapsilosis (20), Cryptococcus neoformans (6), C. krusei (4), C. guilliermondii (3), 及C. lusitaniae (1)，和其他3種臨床較少見的酵母菌。除了這3種少見菌種沒有被鑑定（unidentified）出來，上述8種目標酵母菌皆被正確的鑑定出來，實驗結果顯示敏感度為98.7%。本實驗方法是一個簡單且快速的方法，它能在6小時內鑑定陽性血瓶中約99%的酵母菌菌株。

A multiplex PCR method using one universal and eight species-specific primers was developed to rapidly identify eight species of yeasts commonly found in positive blood cultures. The species-specific primers were designed from the internal transcribed spacer (ITS) regions 1 and 2 of the rRNA gene, whereas the universal primer was located at the 26S rRNA gene. The amplicon sizes of the eight yeast species were as follows: C. lusitaniae (116 bp), C. parapsilosis (126 bp), C. tropicalis (149 bp), C. guilliermondii (185 bp), C. albicans (402 to 403 bp), C. krusei (475 bp), Cryptococcus neoformans (516 bp), and C. glabrata (631 to 632 bp). The PCR products could be effectively separated by gel electrophoresis. By testing 248 clinical isolates of the eight species, a sensitivity of 100% was obtained. However, a specificity of 98.2% was obtained with 168 strains (71 species representing 36 genera) of other pure cultures of yeast and mold tested. The method was then used to analyze 234 positive blood cultures (237 isolates), from which the following species (strain number) were isolated: C. albicans (121), C. tropicalis (50), C. glabrata (29), C. parapsilosis (20), Cryptococcus neoformans (6), C. krusei (4), C. guilliermondii (3), C. lusitaniae (1), and other minor species (3). Except the three minor species, all isolates of the eight target species were identified by the multiplex PCR, resulting in a test sensitivity of 98.7% (234/237). The present method is simple, rapid, and approximately 99% of yeasts in positive blood cultures could be identified within 6 h.

中文摘要 -------------------------------------------------------------------- I
英文摘要 -------------------------------------------------------------------- II
誌謝 -------------------------------------------------------------------------- III
目錄 -------------------------------------------------------------------------- IV
表目錄 ----------------------------------------------------------------------- VI
圖目錄 ----------------------------------------------------------------------- VII
第一章 緒論 ----------------------------------------------------------------- 01
1.1 前言------------------------------------------------------------------- 01
1.2 真菌的特性與分類分類------------------------------------------- 01
1.3 醫學上真菌感染所引起的病症---------------------------------- 04
1.4 酵母菌之院內感染與治療---------------------------------------- 07
1.5 酵母菌之鑑定方法------------------------------------------------- 10
1.6 研究目的與動機---------------------------------------------------- 13
1.7 研究架構------------------------------------------------------------- 15
第二章 實驗方法與材料--------------------------------------------------- 17
2.1 實驗原理------------------------------------------------------------- 17
2.1.1 聚合酶連鎖反應(polymerase chain reaction)---------- 17
2.1.2 瓊脂電泳(AGE) -------------------------------------------- 18
2.1.3 聚丙醯胺凝膠電泳(PAGE)------------------------------- 19
2.1.4 定序分析(DNA sequencing)------------------------------ 19
2.2 實驗材料與方法---------------------------------------------------- 21
2.2.1 實驗藥品----------------------------------------------------- 21
2.2.2 實驗藥品配製----------------------------------------------- 23
2.2.3 實驗設備----------------------------------------------------- 28
2.2.4 真菌和細菌菌種來源-------------------------------------- 31
2.2.5 DNA萃取--------------------------------------------------- 32
2.2.6 內轉錄區（ITS）1和2的放大-------------------------- 35
2.2.7 多套式PCR的引子設計---------------------------------- 36
2.2.8 多套式PCR-------------------------------------------------- 37
2.2.9 血液培養----------------------------------------------------- 38
2.2.10 多套式PCR的偵測極限---------------------------------- 38
2.2.11 膠體電泳----------------------------------------------------- 38
2.2.12 核甘酸序列資料庫登錄號碼----------------------------- 39
第三章 結果------------------------------------------------------------------ 40
3.1 ITS區域序列相似性分析----------------------------------------- 40
3.2 多套式PCR產物的長度------------------------------------------ 40
3.3 多套式PCR的偵測極限------------------------------------------ 41
3.4 以多套式PCR臨床分離株的純培養--------------------------- 41
3.5 其他菌株------------------------------------------------------------- 42
3.6 陽性血瓶中酵母菌之鑑定---------------------------------------- 42
第四章 討論 ----------------------------------------------------------------- 44
參考文獻 --------------------------------------------------------------------- 48

1. Abliz, P., K. Fukushima, K. Takizawa, R. Yang, R. Li, and K. Nishimura. 2002. Identification of the first isolates of Trichosporon asahii var. asahii from disseminated trichosporonosis in China. Diagn. Microbiol. Infect. Dis. 44:17-22.
2. Abu Al-Sound, W., and P. Rådström. 2001. Purification and characterization of PCR-inhibitory components in blood cells. J. Clin. Microbiol. 39:485-493.
3. Abu Al-Sound, W., L. J. Jönsson, and P. Rådström. 2000. Identification and characterization of immunoglobulin G in blood as a major inhibitor of diagnostic PCR. J. Clin. Microbiol. 38:345-350.
4. Ahmad, S., Z. Khan, A. S. Mustafa, and Z. U. Khan. 2002. Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical methods for species identification. J. Clin. Microbiol. 40: 2483-2489.
5. Ainsworth, G. C. 1986. History of medical and veterinary mycology, Cambridge University Press. Cambridge. p. 43—47.
6. Annemarie, B., A. Maurine, L. V. Hall, J. Verhoef, and A. D. Fluit. 2001. Detection of Candida spp. in blood cultures using nucleic acid sequence-based amplification (NASBA). Diagn. Microbiol. infect. Dis. 39:155-160.
7. Aragao, P. A., I. C. Oshiro, E. I. Manrique, C. C. Gomes, L. L. Matsuo, C. Leone, M. L.Moretti-Branchini, and A. S. Levin; IRIS Study Group. 2001. Pichia anomala outbreak in a nursery: exogenous source. Pediatr. Infect. Dis. J. 20:843-848.
8. Averbuch, D., T. Boekhoutt, R. Falk, D. Engelhard, M. Shapiro, C. Block, and I. Polacheck. 2002. Fungemia in a cancer patient caused by fluconazole-resistant Cryptococcus laurentii. Med. Mycol. 40:479-484.
9. Bauters, T. G., M., D. Swinne, V. Stove, and H. J. Nelis. 2003. Detection of single cells of Cryptococcus neoformans in clinical samples by solid-phase cytometry. J. Clin. Microbiol. 2003. 41: 1736-1737.
10. Banerjee, S. N., T. G. Emori, D. H. Culver, R. P. Gaynes, W. R. Jarvis, T. Horan, J. E. Edward, J. Tolson, and W. J. Martine. 1991. Secular trends in nosocomial primary blood-stream infections in the United State, 1980-1989. National Nosocomial Infections Surviellance System. Am. J. Med. 91:86-89.
11. Beck-Sague, C., and W. R. Jarvis. 1993. Secular trends in the epidemiology of nosocomial fungal infections in the United States, 1980-1990. National Nosocomial Infections Surveillance System. J. Infect. Dis. 167:1247-1251.
12. Botelho, A. R., and R. J. Planta. 1994. Specific identification of Candida albicans by hybridization with oligonucleotides derived from ribosomal DNA internal spacers. Yeast 10:709-717.
13. Brandt, M. E., L. E. Benjamin, and G. E. Steinkraus. 2003. Pseudooutbreak of Candida versatilitis fungemia in a microbiology laboratory. Diagn. Microbiol. Infect. Dis. 46:73-75.
14. Chang, H. C., S. N. Leaw, A. H. Huang, T. L. Wu, and T. C. Chang. 2001. Rapid identification of yeasts in positive blood cultures by a multiplex PCR method. J. Clin. Microbiol. 39:3466-3471.
15. Chang, H. C., J. J. Chang, A. H. Huang, and T. C. Chang. 2000. Evaluation of a capacitance method for direct antifungal susceptibility testing of yeasts in positive blood cultures. J. Clin. Microbiol. 38:971-976.
16. Chen, M. L., Y. C. Chen, H. J. Pan, S. C. Chang, L. S. Yang, S. W. Ho, K. T. Luh, W. C. Hsieh, and C. Y. Chuang. 1995. Secular trends in the etiology of nosocomial infection at a teaching hospital in Taiwan, 1981-1994. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi. 28:203-217.
17. Chang, S. E., K. J. Kim, W. S. Lee, J. H. Choi, K. J. Sung, K. C. Moon, and J. K. Koh. 2003. A case of Trichosporon cutaneum folliculitis and septicaemia. Clin. Exp. Dermatol. 28:37-38.
18. Chen, Y. C., J. D. Eisner, M. M. Kattar, S. L. Rassoulian-Barrett, K. Lafe, S. L. Yarfitz, A. P. Limaye, and B. T. Cookson. 2000. Identification of medically important yeasts using PCR-based detection of DNA sequence polymorphisms in the internal transcribed spacer 2 region of the rRNA genes. J. Clin. Microbiol. 38:2302-2310.
19. Chen, Y. C., S. C. Chang, C. C. Sun, L. S. Yang, W. C. Hsieh, K. T. Luh. 1997. Secular trends in the etiology of nosocomial fungal infection at a teaching hospital in Taiwan. Infect Control Hosp. Epidemiol. 18:369-375.
20. Chryssanthou, E., U. Broberger, and B. Petrini. 2001. Malassezia pachydermatis fungaemia in a neonatal intensive care unit. Acta Paediatr. 90:323-327.
21. Chu, C. M., and R. W. Lai. 2002. Malassezia furfur fungaemia in a ventilator-dependent patient without known risk factors. Hong Kong Med. J. 8:212-214.
22. Chung, J. W., B. N. Kim, and Y. S. Kim. 2002. Central venous catheter-related Rhodotorula rubra fungemia. J. Infect. Chemother. 8:109-110.
23. Colombo, A. L., L. S. Dantas, M. L. Abramczyk, M. Cypriano, O. Fischman, A.V. Iazzetti, A. S. Petrilli, and M. P. Selijan. 1997. Rhodotorula glutinis fungemia: a case report and literature review. Braz. J. Infect. Dis. 1:204-207.
24. D'Antonio, D., F. Romano, E. Pontieri, G. Fioritoni, C. Caracciolo, S. Bianchini, P. Olioso, T. Staniscia, R. Sferra, S. Boccia, A. Vetuschi, G. Federico, E. Gaudio, and G. Carruba. 2002. Catheter-related candidemia caused by Candida lipolytica in a patient receiving allogeneic bone marrow transplantation. J. Clin. Microbiol. 40:1381-1386.
25. D'Antonio, D., B. Violante, A. Mazzoni, T. Bonfini, M. A. Capuani, F. D'Aloia, A. Iacone, F. Schioppa, and F. Romano. 1998. A nosocomial cluster of Candida inconspicua infections in patients with hematological malignancies. J. Clin. Microbiol. 36:792-795.
26. de Bernardis, F., R. Lorenzini, R. Verticchio, L. Agatensi, and A. Cassone. 1989. Isolation, acid proteinase secretion, and experimental pathogenicity of Candida parapsilosis from outpatients with vaginitis. J. Clin. Microbiol. 27:2598-2603.
27. de Hoog, G. S., J. Guarro, J. Gene, M. J. Figueras. 2000. Atlas of clinical fungi. 2nd edition. Centraalbureau voor Schimmelcultures. The Netherlands. p. 21-30.
28. Dóczi, I., E. Dósa, E. Hajdú, and E. Nagy. 2002. Aetiology and antifungal susceptibility of yeast bloodstream infections in a Hungarian university hospital between 1996 and 2000. J. Med. Microbiol. 51:677-681.
29. Edwards, M. C., and R. A. Gibbs. 1995. Multiplex PCR, p. 157-171. In C. W. Dieffenbach, and G. S. Dveksler (ed.), PCR primer: a laboratory manual. Cold Spring Harbor Laboratory Press, Plainview, New York.
30. Einsele, H., H. Hebart, G. Goller, J. Loffler, I. Rothenhofer, C. A. Muller, R. A. Bowden, J. V. Burik, D. Engelhard, L. Kanz, and U. Schumacher. 1997. Detection and identification of fungal pathogens in blood by using molecular probes. J. Clin. Microbiol. 35: 1353-1360.
31. Elie, C. M., T. J. Lott, E. Reiss, and C. J. Morrison. 1998. Rapid identification of Candida species with species-specific DNA probes. J. Clin. Microbiol. 36:3260-3265.
32. Elizabeth M. L., 1996. Fundamentals of the fungi. four edition. p. 1-14.
33. Flahaut, M., D. Sanglard, M. Monod, J. Billie, and M. Rossier. 1998. Rapid detection of Candida albicans in clinical samples by DNA amplification of common regions from C. albicans-secreted aspartic proteinase genes. J. Clin. Microbiol. 36: 395-401.
34. Fujita, S. I., Y. Senda, S. Nakaguchi, and T. Hashimoto. 2001. Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains. J. Clin. Microbiol. 39: 3617-3622.
35. Garbino, J., L. Kolarova, P. Rohner, D. Lew, P. Pichna, D. Pittet. 2002. Secular trends of candidemia over 12 years in adult patients at a tertiary care hospital. Medicine. 81:425-433.
36. Guarro, J., M. Nucci, T. Akiti, J. Gene, M.D. Barreiro, and R. T. Goncalves. 2000. Fungemia due to Fusarium sacchari in an immunosuppressed patient. J. Clin. Microbiol. 38:419-421.
37. Hazen, K. C. 1995. New and emerging yeast pathogens. Clin. Microbiol. Rev. 8:462-478.
38. Hendolin, P. H., L. Paulin, P. Koukila-kahkola, V. Anttila, H. Malmberg, M. Richardson, and J. Ylikoski. 2000. Panfungal PCR and multiplex liquid hybridization for detection of fungi in tissue specimens. J. Clin. Microbiol. 38:4186-4192.
39. Henry, T., P. C. Iwen, and S. H. Hinrichs. 2000. Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. J. Clin. Microbiol. 38:1510-1515.
40. Horre, R., S. Gilges, P. Breig, B. Kupfer, G. S. de Hoog, E. Hoekstra, N. Poonwan, and K. P. Schaal. 2001. Case report. Fungaemia due to Penicillium piceum, a member of the Penicillium marneffei complex. Mycoses 44:502-504.
41. Hsueh, P. R., L. J. Teng, P. C. Yang, S. W. Ho, K. T. Luh. 2002. Emergence of nosocomial candidemia at a teaching hospital in Taiwan from 1981 to 2000: increased susceptibility of Candida species to fluconazole. Microb. Drug Resist. 8:311-319.
42. Jaeger, E. E. M., N. M. Carroll, S. Choudhury, A. A. S. Dunlop, H. M. A. Towler, M. M. Matheson, P. Adamson, N. Okhravi, and S. Lightman. 2000. Rapid detection and identification of Candida, Aspergillus, and Fusarium species in ocular samples using nested PCR. J. Clin. Microbiol. 38:2902-2908.
43. Kalenic, S., M. Jandrlic, V. Vegar, N. Zuech, A. Sekulic, and E. Mlinaric-Missoni. 2001. Hansenula anomala outbreak at a surgical intensive care unit: a search for risk factors. Eur. J. Epidemiol. 17:491-496.
44. Kellogg, J. A., J. P. Manzella, and D. A. Bankert. 2000. Frequency of low-level bacteremia in children from birth to fifteen years of age. J. Clin. Microbiol. 38:2181-2185.
45. Krcmery, V., and A. Kunova. 2000. Candida famata fungemia in a cancer patient: case report. J. Chemother. 12:189-90.
46. Kustimur, S., A. Kalkanci, K. Caglar, M. Dizbay, F. Aktas, and T. Sugita. 2002. Nosocomial fungemia due to Trichosporon asteroides: firstly described bloodstream infection. Diagn. Microbiol. Infect. Dis. 43:167-170.
47. Lherm, T., C. Monet, B. Nougiere, M. Soulier, D. Larbi, G. C. Le, D. Caen, and C. Malbrunot. 2002. Seven cases of fungemia with Saccharomyces boulardii in critically ill patients. Intens. Care Med. 28:797-801.
48. Lin, D., L. Wu, M. G. Rinaldi, and P. F. Lehmann. 1995. Three distinct genotypes within Candida parapsilosis from clinical sources. J. Clin. Microbiol. 33:1815-1821.
49. Lo, H. J., Y. A. Ho, and H. Monto. 2001. Factors accounting for misidentification of Candida species. J. Microbiol. Immunol. Infect. 34:171-177.
50. Loeffler, J., N. Henke, H. Hebart, D. Schmidt, L. Hagmeyer, U. Schumacher, and H. Einsele. 2000. Quantification of fungal DNA by using fluorescence resonance energy transfer and the light cycler system. J. Clin. Microbiol. 38: 586-590.
51. Luh, K. T. 1998. Penicillium marneffei fungemia in an AIDS patient: the first case report in Taiwan. Changgeng Yi Xue Za Zhi. 21:362.
52. Luo, G. and T. G. Mitchell. 2002. Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR. J. Clin. Microbiol. 40: 2860-2865.
53. Luzzati, R., G. Amalfitano, L. Lazzarini, F. Soldani, S. Bellino, M. Solbiati, M. C. Danzi, S. Vento, G. Todeschini, C. Vivenza, and E. Concia. 2000. Nosocomial candidemia in non-neutropenic patients at an Italian tertiary care hospital. Eur. J. Clin. Microbiol. Infect. Dis. 19:602-607.
54. Makimura, K., S. Y. Murayama, and H. Yamaguchi. 1994. Detection of a wide range of medically important fungi by the polymerase chain reaction. J. Med. Microbiol. 40:358-364.
55. Martin, C., D. Roberts, M. Van Der Weide, R. Rossau, G. Jannes, T. Smith, and M. Maher. 2000. Development of a PCR-based line probe assay for identification of fungal pathogens. J. Clin. Microbiol. 38:3735-3742.
56. Mathews, C. K., and K. E. van Holde. 1996. Biochemistry, 2nd edition. The Benjamin/Cummings Publishing Company, California.
57. McKelvie, P. A., E. Y. Wong, C L. P. How, and A. J. Hall. 2001. Scedosporium endophthalmitis: two fatal disseminated cases of Scedosporium infection presenting with endophthalmitis. Clin. Experiment. Ophthalmol. 29: 330-334.
58. McMullan, R., J. Xu, J. E. Moore, B. C. Millar, M. J. Walker, S. T. Irwin, J. Price, J. Barr, and S. Hedderwick. 2002. Candida dubliniensis bloodstream infection in patients with gynaecological malignancy. Eur. J. Clin. Microbiol. Infect. Dis. 21:635-636.
59. Morace, G., L Pagano, M. Sanguinetti, B. Posteraro, L. Mele, F. Equitani, G. D’Amore, G. Leone, and G. Fadda. 1999. PCR-restriction enzyme analysis for detection of Candida DNA in blood from febrile patients with hematological malignancies. J. Clin. Microbiol. 37:1871-1875.
60. Morace, G., M. Sanguinetti, B. Posteraro, G. Lo. Cascio, and G Fadda. 1997. Identification of various medically important Candida species in clinical specimens by PCR-restriction enzyme analysis. J. Clin. Microbiol. 35: 667-672.
61. Nedret, K. A., F. Erdem, and T. Patiroglu. 2002. Case report. Acremonium falciforme fungemia in a patient with acute leukaemia. Mycoses 45:202-203.
62. Neumeister, B., M. Rockemann, and R. Marre. 1992. Fungemia due to Candida pelliculosa in a case of acute pancreatitis. Mycoses 35:309-310.
63. Nucci, M., T. Akiti, G. Barreiros, F. Silveira, and S. G. Revankar, B. L. Wickes, D. A. Sutton, and T. F. Patterson. 2002. Nosocomial outbreak of Exophiala jeanselmei fungemia associated with contamination of hospital water. Clin. Infect. Dis. 34:1475-1480.
64. Olive, D. M., and P. Bean. 1999. Minireview: principles and applications of methods for DNA-based typing of microbial organisms. J. Clin. Microbiol. 37:1661-1667.
65. Pfaller, M. A., and D. J. Diekema. 2002. Role of sentinel surveillance of candidemia: trends in species distribution and antifungal susceptibility. J. Clin. Microbiol. 40:3551-3557.
66. Pfaller, M. A., D. J. Diekema, R. N. Jones, S. A. Messer, R. J. Hollis, and the SENTRY Participants Group. 2002. Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adult patients with bloodstream infections: SENTRY antimicrobial surveillance program, 1997 to 2000. J. Clin. Microbiol. 40:852-856.
67. Pfaller, M. A., S. A. Messer, R. J. Hollis, R. N. Jones, G. V. Doern, M. E. Brandt, and R. A. Hajjeh. 1999. Trends in species distribution and susceptibility to fluconazole among blood stream isolates of Candida species in the United States. Mycology 33:217-222.
68. Pfaller, M. A., R. N. Jones, S. A. Messer, M. B. Edmond, R. P. Wenzel, and the SCOPE participant group. 1998. National surveillance of nosocomial blood stream infection due to species of Candida other than Candida albicans: frequency of occurrence and antifungal susceptibility in the SCOPE program. Diagn. Microbiol. Infect. Dis. 30:121-129.
69. Peltroche-Llacsahuanga, H., E. Manegold, G. Kroll, and G. Haase. 2000. Case report. Pathohistological findings in a clinical case of disseminated infection with Fusarium oxysporum. Mycoses 43:367-372.
70. Perapoch, J., A. M. Planes, A. Querol, V. Lopez, I. Martinez-Bendayan, R. Tormo, F. Fernandez, G. Peguero, and S. Salcedo. 2000. Fungemia with Saccharomyces cerevisiae in two newborns, only one of whom had been treated with ultra-levura. Eur. J. Clin. Microbiol. Infect. Dis. 19:468-70.
71. Pittet, D., and R. P. Wenzel. 1995. Nosocomial bloodstream infections. Arch. of Intern. Med. 155:1177-1184.
72. Quindos, G., R. S. Millan, R. Robert, C. Bernard and J. Ponton. 1997. Evaluation of bichro-latex albicans, a new method for rapid identification of Candida albicans. J. Clin. Microbiol. 35:1263-1265.
73. Raj, R., and A. E. Frost. 2002. Scedosporium apiospermum fungemia in a lung transplant recipient. Chest 121:1714-1716.
74. Reimer, L. G., M. L. Wilson, and M. P. Weinstein. 1997. Update on detection of bacteremia and fungemia. Clin. Microbiol. Rev. 10:444-465.
75. Reiss, E., K. Tanaka, G. Bruker, V. Chazalet, D. Coleman, J. P. Debeaupuis, R. Hanazawa, J. P. Latge, J. Lortholary, K. Makimura, C. J. Morrison, S. Y. Murayama, S. Naoe, S. Paris, J. Sarfati, K. Shibuya, D. Sullivan, K. Uchida, and H. Yamaguchi. 1998. Molecular diagnosis and epidemiology of fungal infections. Med. Mycol. Suppl. 1:249-257.
76. Rex, J. H., J. E. Bennett, A. M. Sugar, P. G. Pappas, C. M. van der Horst, J. E. Edwards, R. G. Washburn, W. M. Scheld, A. W. Karchmer, A. P. Dine, M. J. Levenstein, and C. D. Webb. 1994. A randomized trial comparing fluconazole with amphotericin B for the treatment of candidemia in patients without neutropenia. N. Engl. J. Med. 331: 1325-1330.
77. Rigby, S., G. W. Procop, G. Haase, D. Wilson, G. Hall, C. Kurtzman, K. Oliveira, S. V. Oy, J. J. Hyldig-Nielsen, J. Coull, and H. Stender. 2002. Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles. J. Clin. Microbiol. 2002 40: 2182-2186.
78. Rodero, L., M. Cuenca-Estrella, S. Cordoba, P. Cahn, G. Davel, S. Kaufman, L. Guelfand, and J. L. Rodriguez-Tudela. 2002. Transient fungemia caused by an amphotericin B-resistant isolate of Candida haemulonii. J. Clin. Microbiol. 40:2266-2269.
79. Rodriguez-Villalobos, H., A. Georgala, H. Beguin, C. Heymans, G. Pye, F. Crokaert, and M. Aoun. 2003. Disseminated infection due to Cylindrocarpon (Fusarium) lichenicola in a neutropenic patient with acute leukaemia: report of a case and review of the literature. Eur. J. Clin. Microbiol. Infect. Dis. 22:62-65.
80. Saiki R. K., S. Scharf, F. Faloona, K. B. Mullis, G. T. Horn, H. A. Erlich, and N. Arnheim. 1992. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. 1985. Biotechnology 24:476-480.
81. Sambrook, J. and D. W. Russell, 2001. Molecular Cloning. Third edition 1: 5.1.
82. Sambrook, J. and D. W. Russell, 2001. Molecular Cloning. Third edition 2: 8.1.
83. Sambrook, J. and D. W. Russell, 2001. Molecular Cloning. Third edition 2: 12.1.
84. Samonis, G., M. Anatoliotaki, H. Apostolakou, S. Maraki, D. Mavroudis, and V. Georgoulias. 2001. Transient fungemia due to Rhodotorula rubra in a cancer patient: case report and review of the literature. Infection 29:173-176.
85. Shin, D.H., J. H. Park, J. H. Shin, S. P. Suh, D. W. Ryang, and S. J. Kim. 2003. Pichia ohmeri fungemia associated with phlebitis: successful treatment with amphotericin B. J. Infect. Chemother. 9:88-89.
86. Shin, J. H., F. S. Nolte, and C. J. Morrison. 1997. Rapid identification of Candida species in blood cultures by a clinical useful PCR method. J. Clin. Microbiol. 35:1454-1459.
87. Simarro, E., F. Marin, A. Morales, E. Sanz, J. Perez, and J. Ruiz. 2001. Fungemia due to Scedosporium prolificans: a description of two cases with fatal outcome. Clin. Microbiol. Infect. 7:645-647.
88. St-Germain, G., and R. Summerbell. 1996. Identifying filamentous fungi. p. 1-8. Star, California.3.
89. Tan, C. S., R. G. Wintermans, G. S. de Hoog, H. W. Engel, and E. P. Ijzerman. 1992. Verschuivingen in het soortenspectrum van mycosen in Nederland in het tijdvak 1970-1990. Ned. Tijdschr. Geneeskd. 136:631-637.
90. Turenne, C. Y., S. E. Sanche, D. J. Hoban, J. A. Karlowsky, and A. M. Kabani. 1999. Rapid identification of fungi by using the ITS2 genetic region and an automated fluorescent capillary electrophoresis system. J. Clin. Microbiol. 37:1846-1851.
91. Wahyuningsih, R., H. J. Freisleben, H. G. Sonntag, and P. Schnitzler. 2000. Simple and rapid detection of Candida albicans DNA in serum by PCR for diagnosis of invasive candidasis. J. Clin. Microbiol. 38:3016-3021.
92. Wang, H. Y., and J. L. Lin. 1999. Trichosporon beigelii fungaemia in a patient with haemodialysis. Nephrol Dial Transplant. 14:2017-2018.
93. Warren, N. G., and K. C. Hazen. 1999. Candida cryptococcus, and other yeasts of medical importance. p. 1184-1199. In P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H. Yolken (ed.), Manual of clinical microbiology, 7th ed. American Society for Microbiology, Washington, D.C.
94. Warris, A., F. Wesenberg, P. Gaustad, P. E. Verweij, and T. G. Abrahamsen. 2000. Acremonium strictum fungaemia in a paediatric patient with acute leukaemia. Scand. J. Infect. Dis. 32 :442-444.
95. Weinstein, M. P., M. L. Towns, S. M. Quartey, S. Mirrett, L. G. Reimer, G. Parmigiani, and L. B. Reller. 1997. The clinical significance of positive blood cultures in the 1990s: a prospective comprehensive evaluation of microbiology, epidemiology, and outcome of bacteremia and fungemia in adults. Clin. Infect. Dis. 24:584-602.
96. White, T. J., T. Bruns, S. Lee, and J. Taylor. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, p. 315-322. In M. A. Innis, D. H. Gefland, J. J. Sninsky, and T. J. White (ed.), PCR protocols: a guide to methods and applications. Academic Press, San Diego, Calif.
97. Williams, D. W., M. J. Wilson, M. A. Lewis, and A. J. Potts. 1995. Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA. J. Clin. Microbiol. 33: 2476-2479.
98. 林春珠、盧柏樑、黃高彬，2000，某醫學中心1985年至1996年院內感染致病菌之變遷與分析，院內感染控制雜誌，10：301-312。
99. 蔡文城，2000，“臨床微生物學”，九州圖書，台北，p. 2-4。