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研究生:陳俊男
研究生(外文):Jin-Nan Chen
論文名稱:北台灣志賀氏宋內菌分離株之抗藥性機制與分子流行病學研究
論文名稱(外文):Antibiotics Resistance Mechanism and Molecular Epidemiology of Shigella Sonnei Isolated from Northern Taiwan
指導教授:胡 文 熙
指導教授(外文):Wensi S. Hu
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:69
中文關鍵詞:志賀氏宋內桿菌脈衝式電泳抗藥基因質體
外文關鍵詞:Shigella sonneiPFGEantibitics resistant geneplasmid
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中文摘要:
本論文乃分析疾病管制局於2001年從北台灣三個山地村落所分離出志賀氏宋內桿菌, 其中新竹地區20株, 桃園地區22株,與宜蘭地區36株, 共78株散發菌株。分別以脈衝式電泳(PFGE)、質體型式 (PPA)等技術進行研究。此78株菌以脈衝氏電泳法分型可將分成7型,分別命名為A-G, 其中以A型最多, 約佔84.6%。質體型式分析共分成10型,分別命名為(P1-P10) 其中以P1最多,約佔有75.6%。 經統計分析結果,其DI(discrimination index) 值分別為0.264與0.421, 表示三地菌株間差異性不大,因此推論可能有一優勢菌株(endemic strain)互相傳遞於三個村落之間。 接著我們探討這些菌株的12種藥物敏感性測試, 發現大部分菌株(89.7%)都帶有4抗藥性{tetracycline (TET)、streptomycin (STR)、trimethoprim-sulfamethoxazale (SXT)、nalidixic acid (NAL)}此外本文更進一步研究探討TET、STR、SXT抗藥性來源與藥物敏感性測試之關聯。並且從抗藥性來源研究上發現,TET、 STR、 sulfamethoxazale的抗藥性是來自於廣泛散佈在此78株菌內的一個8.5kb質體 (暫命名為pR), 並以PCR確定質體(pR)上帶有tetA、strA、sulII三種抗藥基因。有別於以往多重抗藥基因質體均為的大分子量(大於20kb), 這個帶多重抗藥基因的小分子量質體是一個全新的發現。 除了上述三種抗藥基因, 此外志賀氏宋內桿菌genomic DNA所帶有的dhfr Ia (trimethoprim抗藥基因), 造成此三地菌株對此三種抗生素大規模的抗藥性包含抗TET(91%) 、STR (91%)、 SXT(89%)。另外也在新竹與宜蘭也發現抗多重β-lactams(cephalothin, cefamandole, ceftriaxone, ampicillin)的菌株存在, 顯示出extended spectrum β-lactamase (ESBL)正逐漸出現於台灣志賀氏宋內桿菌中。 進一步以PCR與DNA sequencing後, 發現新竹地區β-lactamase基因為 CTXM-14, 而宜蘭地區為cmy-2。 這些抗藥基因研究有助於臨床治療痢疾時用藥上的參考。
英文摘要:
Seventy-eight isolates of Shigella sonnei were obtained from sporadic clinical infection in northern part of Taiwan, 20 isolates from Hsin-chu county, 22 isolates from Tao-yuan county, and 36 islates from I-lan county. Pulsed-field gel electrophoresis (PFGE) and plasmid profile analysis(PPA) were applied for epidemiological study. Seven types (A, B, C, D, E, F, G) were classified based on PFGE, and type A is the majority (84.5%). On the other hands, ten types (P1, P2, P3, P4, P5, P6, P7, P8, P9, and P10) were also classified based on PPA, and type P1 is the majority (75.6%). Using statistical analysis, the DI value of PEGE and PPA are 0.263 and 0.421, respectively, which indicated that these 78 isolates were closely related. Therefore, we predicted that a possible endemic strain existed in these counties. A total of 12 antibiotics were used for susceptibility test of these 78 isolates by using Kirby-Bauer disk diffusion test. The result showed that 91%(71/78) of those 78 isolates was resistant at least to streptomycin (STR), trimethoprim-sulfamethozale (SXT), tetracycline (TET), and nalidixic acid (NAL). A novel 8.5 kb plasmid (PR), carrying STR and TET resistance, was confirmed by transformation assay. Furthermore, tetA, strA, sulII resistance genes were identified by polymerase chain reaction (PCR) assay. This is different from previously reports that a large plasmid mediated STR and TET resistance. For the SXT resistance, dhfr Ia and sul II genes were identified in these resistant isolates. Among these 78 isolates, there are 8 isolates from Hsin-chu county and 3 isolates from I-lan county which having extended spectrum ß-lactamase (ESBL) resistance. CTXM-14 resistant gene in 7 strains of Hsin-chu county and cmy-2 gene in 3 isolates of I-lan county were identified, respectively. Identification of these antibiotics-resistant genes is to provide update information for clinician to treat diarrhea caused by S. sonnei.
目錄
致 謝--------------------------------------------------------3
中文摘要--------------------------------------------------------4
英文摘要--------------------------------------------------------6
緒 論--------------------------------------------------------8
材料方法--------------------------------------------------------18
結 果--------------------------------------------------------29
討 論--------------------------------------------------------37
參考文獻--------------------------------------------------------41
圖片說明--------------------------------------------------------48
表 格--------------------------------------------------------61
附 錄--------------------------------------------------------66
1. 潘子明. 痢疾之流行趨勢與預防. 疫情報導 1996; 12: 212-19.
2. 林建生, 王添貴, 蔡金來, 何淑英,李智隆. 台灣桃園地區宋內氏志賀氏桿菌感染事件之關聯性分析. 疫情報導 1999. 17(4):169-177.
3. 邱乾順, 沈玉梅, 楊麗珠, 魏孝倫, 廖采苓. 南投縣桿菌性痢疾流行概況-1995至1999年. 疫情報導. 1999. 17(7):341-58.
4. 林建生, 王添貴, 蔡金來, 何淑英, 潘子明. 脈場膠電泳和質體輪廓分析台灣地區1995-1996大規模細菌性痢疾桿菌感染事件. 台灣醫學. 1997. 2: 152-8.
5. Olive DM, Bean P. Principles and applications of methods for DNA-based typing of microbial organisms.J Clin Microbiol. 1999 Jun;37(6):1661-9. Review.
6. Chiou CS, Hsu WB, Wei HL, Chen JH Molecular epidemiology of a Shigella flexneri outbreak in a mountainous township in Taiwan, Republic of China.J Clin Microbiol. 2001 Mar;39(3):1048-56.
7. Liu PY, Lau YJ, Hu BS, Shyr JM, Shi ZY, Tsai WS, Lin YH, Tseng CY. Analysis of clonal relationships among isolates of Shigella sonnei by different molecular typing methods.J Clin Microbiol. 1995 Jul;33(7):1779-83.
8. Litwin CM, Storm AL, Chipowsky S, Ryan KJ. Molecular epidemiology of Shigella infections: plasmid profiles, serotype correlation, and restriction endonuclease analysis.J Clin Microbiol. 1991 Jan;29(1):104-8.
9. Wong HC, Lu KT, Pan TM, Lee CL, Shih DY. Subspecies typing of Vibrio parahaemolyticus by pulsed-field gel electrophoresis.
J Clin Microbiol. 1996 Jun;34(6):1535-9.
10. Liebisch B, Schwarz S. Molecular typing of Salmonella enterica subsp. enterica serovar Enteritidis isolates.J Med Microbiol. 1996 Jan;44(1):52-9.
11. Goering RV. Molecular epidemiology of nosocomial infection: analysis of chromosomal restriction fragment patterns by pulsed-field gel electrophoresis.Infect Control Hosp Epidemiol. 1993 Oct;14(10):595-600. Review.
12. Rajakumar K, Bulach D, Davies J, Ambrose L, Sasakawa C, Adler B. Identification of a chromosomal Shigella flexneri multi-antibiotic resistance locus which shares sequence and organizational similarity with the resistance region of the plasmid NR1.
Plasmid. 1997;37(3):159-68.
13. Grohmann E, Muth G, Espinosa M. Conjugative plasmid transfer in gram-positive bacteria.Microbiol Mol Biol Rev. 2003 Jun;67(2):277-301, table of contents.
14. Adeleye IA. Conjugal transferability of multiple antibiotic resistance in three genera of Enterobacteriaceae in Nigeria.J Diarrhoeal Dis Res. 1992 Jun;10(2):93-6.
15. Ling JM, Shaw PC, Kam KM, Cheng AF, French GL. Molecular studies of plasmids of multiply-resistant Shigella spp. in Hong Kong.
Epidemiol Infect. 1993 Jun;110(3):437-46.
16. McIver CJ, White PA, Jones LA, Karagiannis T, Harkness J, Marriott D, Rawlinson WD. Epidemic strains of Shigella sonnei biotype g carrying integrons.J Clin Microbiol. 2002 Apr;40(4):1538-40.
17. Recchia GD, Hall RM. Gene cassettes: a new class of mobile element.Microbiology. 1995 Dec;141 ( Pt 12):3015-27. Review.
18. Ploy MC, Denis F, Courvalin P, Lambert T. Molecular characterization of integrons in Acinetobacter baumannii: description of a hybrid class 2 integron.Antimicrob Agents Chemother. 2000 Oct;44(10):2684-8
19. White PA, McIver CJ, Deng Y, Rawlinson WD. Characterisation of two new gene cassettes, aadA5 and dfrA17.FEMS Microbiol Lett. 2000 Jan 15;182(2):265-9.
20. Sundstrom L, Skold O. The dhfrI trimethoprim resistance gene of Tn7 can be found at specific sites in other genetic surroundings.
Antimicrob Agents Chemother. 1990 Apr;34(4):642-50.
21. Peter A. White, William D. Randwick. 2001. Current status of and DHFR gene cassette families. J Antimicrob Chemother 47:495-496.
22. Roberts MC. Tetracycline resistance determinants: mechanisms of action, regulation of expression, genetic mobility, and distribution.
FEMS Microbiol Rev. 1996 Oct;19(1):1-24. Review.
23. Massova I, Mobashery S. Kinship and diversification of bacterial penicillin-binding proteins and beta-lactamases.Antimicrob Agents Chemother. 1998 Jan;42(1):1-17. Review
24. Hardt K, Joris B, Lepage S, Brasseur R, Lampen JO, Frere JM, Fink AL, Ghuysen JM. The penicillin sensory transducer, BlaR, involved in the inducibility of beta-lactamase synthesis in Bacillus licheniformis is embedded in the plasma membrane via a four-alpha-helix bundle.Mol Microbiol. 1997 Mar;23(5):935-44.
25. Kuwahara-Arai K, Kondo N, Hori S, Tateda-Suzuki E, Hiramatsu K. Suppression of methicillin resistance in a mecA-containing pre-methicillin-resistant Staphylococcus aureus strain is caused by the mecI-mediated repression of PBP 2'' production.Antimicrob Agents Chemother. 1996 Dec;40(12):2680-5.
26. Bush K, Jacoby GA, Medeiros AA. A functional classification scheme for beta-lactamases and its correlation with molecular structure.Antimicrob Agents Chemother. 1995 Jun;39(6):1211-33. Review.
27. 賴美珠. 產生超廣效性ß-lactamase的分類原理、測定方式及抗藥性問題. 醫檢會報. 2002.17(3): P27-37.
28. Sirot D, Chanal C, Bonnet R, De Champs C, Bret L. Inhibitor -resistant TEM-33 beta-lactamase in a Shigella sonnei isolate.
Antimicrob Agents Chemother. 2001 Jul;45(7):2179-80
29. Sabate M, Tarrago R, Navarro F, Miro E, Verges C, Barbe J, Prats G. Cloning and sequence of the gene encoding a novel cefotaxime -hydrolyzing beta-lactamase (CTX-M-9) from Escherichia coli in Spain.Antimicrob Agents Chemother. 2000 Jul;44(7):1970-3.
30. Pai H, Choi EH, Lee HJ, Hong JY, Jacoby GA. Identification of CTX-M-14 extended-spectrum beta-lactamase in clinical isolates of Shigella sonnei, Escherichia coli, and Klebsiella pneumoniae in Korea.
J Clin Microbiol. 2001 Oct;39(10):3747-9.
31. Rasheed JK, Jay C, Metchock B, Berkowitz F, Weigel L, Crellin J, Steward C, Hill B, Medeiros AA, Tenover FC. Evolution of extended-spectrum beta-lactam resistance (SHV-8) in a strain of Escherichia coli during multiple episodes of bacteremia.
Antimicrob Agents Chemother. 1997 Mar;41(3):647-53.
32. Chang FY, Siu LK, Fung CP, Huang MH, Ho M. Diversity of SHV and TEM beta-lactamases in Klebsiella pneumoniae: gene evolution in Northern Taiwan and two novel beta-lactamases, SHV-25 and SHV-26.
Antimicrob Agents Chemother. 2001 Sep;45(9):2407-13.
33. Siu LK, Lo JY, Yuen KY, Chau PY, Ng MH, Ho PL. beta-lactamases in Shigella flexneri isolates from Hong Kong and Shanghai and a novel OXA-1-like beta-lactamase, OXA-30.Antimicrob Agents Chemother. 2000 Aug;44(8):2034-8.
34. Zhao S, White DG, McDermott PF, Friedman S, English L, Ayers S, Meng J, Maurer JJ, Holland R, Walker RD. Identification and expression of cephamycinase bla(CMY) genes in Escherichia coli and Salmonella isolates from food animals and ground meat.
Antimicrob Agents Chemother. 2001 Dec;45(12):3647-50.
35. Sanchez-Pescador R, Brown JT, Roberts M, Urdea MS. Homology of the TetM with translational elongation factors: implications for potential modes of tetM-conferred tetracycline resistance.
Nucleic Acids Res. 1988 Feb 11;16(3):1218.
36. Sheridan RP, Chopra I. Origin of tetracycline efflux proteins: conclusions from nucleotide sequence analysis.Mol Microbiol. 1991 Apr;5(4):895-900.
37. Shaw KJ, Rather PN, Hare RS, Miller GH. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes.
Microbiol Rev. 1993 Mar;57(1):138-63. Review.
38. Mingeot-Leclercq MP, Glupczynski Y, Tulkens PM. Aminoglycosides: activity and resistance.Antimicrob Agents Chemother. 1999 Apr;43(4):727-37. Review.
39. Shaw PC, Liang AC, Kam KM, Ling JM. Presence of strA-strB gene within a streptomycin-resistance operon in a clinical isolate of Shigella flexneri.Pathology. 1996 Nov;28(4):356-8.
40. Sundin GW. Distinct recent lineages of the strA- strB streptomycin-resistance genes in clinical and environmental bacteria.
Curr Microbiol. 2002 Jul;45(1):63-9.
41. Peter A. White, William D. Randwick. Current status of and DHFR gene cassette families. J Antimicrob Chemother 2001 47:495-496.
42. Sunde M, Fossum K, Solberg A, Sorum H. Antibiotic resistance in Escherichia coli of the normal intestinal flora of swine.Microb Drug Resist. 1998 Winter; 4(4): 289-99.
43. Pentti Huovinen, Lars Sunstrom, Gote Swedberg, Ola skold. 1995. Trimethoprim and sulfonamide resistance. Antimicrob. Agents Chemother. 39(2): P279-289.
44. P. L. Winokur, A. Brueggemann, D.L. Hoffmann, M.A. Pfaller, G.. V. Doern. 2000.Animal and human multidrug-resistant, cephalosporin-resistant Salmonalla Isolates expressing a plasmid-mediated CMY-2 Ampc ß-lactamase. Antimicrob. Agents Chemother. 44(10): P2777-2783.
45. Corinna Kehrenberg, Stefan Schwarz. 2001. Occurremce and linkage of genes coding for resistance to sulfonamides, streptomycin and chloramphenical in bacteria of genera Pasteurella and Mannheimia. FEMS microbio. Lett. 205: P283-290.
46. Gilliane Guillaume, Dirk Verbrugge, Marie-Lousie Chasseur-Libotte, William Moens, Jean-Marc Collard. 2000. PCR typing of tetracycline resistance determination (Tet A-E) in Salmonella enterica serotype Hadar and in the microbial community of activated sludges from hospital and urban wastewater treatment facilities in Belgium. FEMS microbio. Ecology. 32: 77-85.
47. Paul. Hunter, Michael A. Gaston. 1988. Numerical index of the discriminatory ability of typing system: an application of Simposon’s index of diversity. JCM. 26(11): P2465-2466.
48. Fred C. Tenover, Robert D. Arbeit, Richard V. Georing, Patricia A. Mickelsen, Barbara E. Murray, David H. Persing, Bla swaminathan. 1995. Interpreting chromosomal DNA restriction pattern produced by pulsed-field gel electrophoresis: Criteria for bacterial strain typing. JCM. 33(9): P2233-2239.
49. Fred C. Tenover, Robert D. Arbeit. 1997. How to select and interpret molecular strain typing methods for epidemiological studies of bacterial infection: A review for healthcare epidemiologists. Infect Control Hosp Epidemiol. 18: 426- 439.
50. T. J. Barrett, H. Lior, J. H. Green, R. Khakhria, J. G. Wells, B. P. Bell, K. D. Greene, J. Lewis, P. M. Griffin. 1994. Laboratory investigation of multistate food-borne outbreak of Escherichia coli O157:H7 by using pulsed-field gel electrophoresis and phage typing. J. clin. Microbial. 32(12): 3013-3017.
51. Perez-Perez FJ, Hanson ND. Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR.
J Clin Microbiol. 2002 Jun;40(6):2153-62.
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