臺灣博碩士論文加值系統

到目前為止，並沒有任何一個研究是利用新的牙周病分類準則來探討伴放線桿菌五種血清型態與牙周疾病分類之關係。本研究的第一部份是要利用舊的牙周病分類與新的牙周病分類，來探討從北美地區不同型態之牙周病病患的口腔中所取得之牙齦下牙菌斑可培養出的伴放線桿菌，其五種血清型態的分佈頻率；並研究血清型態與年齡或臨床診斷間有無相關性存在。第二部份是探討在台灣地區年輕族群（35歲以下）中患有侵犯性牙周病、慢性牙周病之患者與牙周健康者，其伴放線桿菌血清型態在牙齦下牙菌斑樣本出現之盛行率及比例。在第一部份本研究總共收集來自115位可培養出伴放線桿菌的345菌種，病患之平均年齡為38.0±18.3歲，其中女性佔59%。根據新的牙周病分類準則，33位屬於侵犯性牙周病患者、82位屬於慢性牙周病患者；若根據舊的牙周病分類準則，則6位屬於青春前期牙周病、12位屬於局部性年輕型牙周病、15位屬於年輕後期型牙周病、28位屬於頑強型牙周病、54位屬於成年型牙周病。在第二部份本研究總共收集221個牙齦下牙菌斑樣本，其中171個來自牙周病患（70位為侵犯性牙周病、101位為慢性牙周病），病患平均年齡為25.0±8.2歲；50個來自牙周健康者，平均年齡為18.4±9.5歲。以間接免疫螢光分析法，利用對伴放線桿菌菌種ATCC 29523（血清型態a）、ATCC 43728（血清型態b）、ATCC 33384（血清型態c）、IDH 781（血清型態d）和IDH 1705（血清型態e）有血清型態專一性的多株抗血清來決定伴放線桿菌之血清型態。在第一部份以Chi square檢定檢視血清型態b在不同診斷與年齡分組間之關係。結果發現大部分之病患為單一血清型態，其中22位為血清型態a、44位為血清型態b、30位為血清型態c、1位為血清型態d、3位為血清型態e。11位病患同時具有兩種不同的血清型態，2位病患同時具有三種不同的血清型態，另有2位病患之血清型態無法被鑑定。雖然血清型態b與年齡之相關性不大，但血清型態b仍是18歲以下之小孩子以及年齡介於19歲至35歲之間的年輕成年人病患最主要之血清型態。血清型態d和血清型態e不會出現在年齡小於35歲的病患，但在62位年齡大於35歲之成年人病患，則分別可發現1個血清型態d和3個血清型態e。血清型態b為侵犯性牙周病患最主要之血清型態(60.61%)且血清型態b在侵犯性牙周病患出現之頻率明顯高於慢性牙周病患(P<0.01)。血清型態d和血清型態e則不會出現在任一位侵犯性牙周病之病患。血清型態a (20.73%)、血清型態b (29.27%)與血清型態c (28.05%)在慢性牙周病患之分佈頻率差不多。血清型態b為青春前期牙周病患(50.00%)、局部性年輕型牙周病患(66.67%)與年輕後期型牙周病患(60.00%)最主要之血清型態。在第二部份以單因子變異數分析(ANOVA)計算伴放線桿菌在各種診斷分組之盛行率及比例。結果發現伴放線桿菌之盛行率在侵犯性牙周病患為84.3%、在慢性牙周病患為60.4%、在牙周健康者為64.0%。伴放線桿菌之比例在牙周健康者（0.55±0.08%）明顯低於侵犯性牙周病患（1.05±0.15%）（P<0.05）；而在慢性牙周病患，其伴放線桿菌之比例也明顯低於侵犯性牙周病患(P<0.05)。血清型態b之比例在侵犯性牙周病患（0.66±0.14%）與慢性牙周病患（0.43±0.11%）均明顯高於牙周健康者（0.02±0.01%）(P<0.05)。血清型態c之比例在牙周健康者（0.46±0.08%）顯著高於慢性牙周病患（0.15±0.04%）(P<0.01)。至於血清型態a在三種不同牙周狀況下之比例則沒有任何之顯著差異存在。
綜合兩部分之研究結果可以發現，在北美地區伴放線桿菌血清型態b為侵犯性牙周病患最主要之血清型態，在慢性牙周病患則血清型態b與血清型態c之分佈頻率差不多。在台灣地區年輕族群中，血清型態b為侵犯性牙周病患與慢性牙周病患最主要之血清型態，而血清型態c則較常出現於牙周健康者。

To our knowledge, the association of the five serotypes of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) to the new diagnostic classification scheme described by the American Academy of Periodontology in 1999 has not yet been described. The purpose of this part I study was to characterize the frequencies of the five serotypes of A. actinomycetemcomitans in A. actinomycetemcomitans isolates from various forms of periodontitis using both old and new diagnostic classifications and to determine the relationships between serotype and age and clinical diagnosis. The purpose of this part II study was to investigate prevalence and proportions of A. actinomycetemcomitans serotypes in subgingival plaque samples from a young Taiwanese population with aggressive periodontitis, chronic periodontitis and no periodontal disease. A total of 345 A. actinomycetemcomitans isolates from 115 A. actinomycetemcomitans culture positive subjects (mean age 38.0 ± 18.3 years, 59% female) were collected from part I study. Based on the new classifications, 33 subjects had aggressive periodontitis and 82 chronic periodontitis. According to old classifications, there were six pre-pubertal periodontitis (PPP), 12 localized juvenile periodontitis (LJP), 15 post-localized juvenile periodontitis (PLJP), 28 refractory periodontitis (Ref-P), and 54 adult periodontitis (AP) cases. A total of 221 subgingival plaque samples from 171 diseased subjects (70 had aggressive periodontitis, and 101 had chronic periodontitis) (mean age 25.0 ± 8.2 yr) and 50 periodontally healthy subjects (mean age 18.4 ± 9.5 yr) were screened for A. actinomycetemcomitans from part II study. Serotypes of A. actinomycetemcomitans were determined by an indirect immunofluorescence assay using serotype-specific polyclonal antisera to A. actinomycetemcomitans strains ATCC 29523, ATCC 43728, ATCC 33384, IDH 781 and IDH 1705 (serotype a, b, c, d, and e, respectively). Proportions of serotype b were examined between different diagnostic and age groups with a Chi square test. Most subjects (n = 100, 86.96%) were infected with a single serotype (22 serotype a, 44 serotype b, 30 serotype c, 1 serotype d and 3 serotype e). There were 11 subjects (9.57%) with two serotypes and two subjects (1.74%) with 3 serotypes. Two individuals had isolates lacking any detectable serotype antigen. Serotype b was the predominant serotype in children under 18 years of age and young adults between 19 to 35 years, although serotype b status was not significantly associated with age. Serotypes d and e were not found in patients under 35 years old. In 62 adult patients, one subject had serotype d and three had serotype e. Serotype b was the most common serotype in aggressive periodontitis (60.61%). The proportion of cases with serotype b was significantly higher in aggressive periodontitis compared to chronic periodontitis (P<0.05). Other serotypes were not significantly associated with new diagnostic categories. Serotypes d and e were not detected in aggressive periodontitis. The results of this part II study show that proportions of serotype b of A. actinomycetemcomitans are significantly greater in culture-positive patients with aggressive periodontitis than those with chronic periodontitis. Prevalence (% of positive samples) of A. actinomycetemcomitans was 84.3% in aggressive periodontitis, 60.4% in chronic periodontitis, and 64.0% in periodontally healthy subjects. Proportions of A. actinomycetemcomitans (mean percentage per total bacteria) in periodontally healthy subjects were significantly lower than in aggressive periodontitis subjects. The proportion of serotype b in subjects with aggressive periodontitis and subjects with chronic periodontitis were significantly greater than that in periodontally healthy subjects. The proportion of serotype c in periodontally healthy subjects was much higher than that in chronic periodontitis subjects. The results of this study suggest that prevalence and proportions of A. actinomycetemcomitans are significantly greater in patients with aggressive periodontitis than in those with chronic periodontitis. Serotype b is the predominant serotype of A. actinomycetemcomitans in patients with diseased periodontal conditions. Serotype c is a more common serotype detected in periodontally healthy subjects.

目錄……………………………………………………………………1
圖、表目錄……………………………………………………………3
壹、中文摘要…………………………………………………………5
貳、英文摘要…………………………………………………………8
參、前言………………………………………………………………12
肆、材料及方法………………………………………………………15
一、診斷分組及定義……………………………………………15
二、樣本收集……………………………………………………15
三、取樣、培養及玻片製備……………………………………16
四、抗血清製備…………………………………………………18
五、抗血清的專一性……………………………………………19
六、間接免疫螢光分析法………………………………………19
伍、結果………………………………………………………………22
陸、討論………………………………………………………………26
一、血清型態的分佈頻率與各種不同型態牙周病之關係……26
二、單一血清型態或混合血清型態感染………………………28
三、血清型態的分布比例………………………………………29
四、無法被鑑定的伴放線桿菌血清型態………………………30
五、血清型態分布與年齡之關係………………………………31
六、伴放線桿菌的盛行率與牙周狀況之關係…………………32
七、血清型態與牙周狀況之關係………………………………33
柒、結論………………………………………………………………35
捌、參考文獻…………………………………………………………36
玖、圖片………………………………………………………………46
拾、表格………………………………………………………………57
拾壹、附錄……………………………………………………………71

1. Socransky S, Haffajee A.D. The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol 1992; 63: 322-331.
2. Lai C, Listgarten M. Comparative ultrastructure of leukotoxic and non-leukotoxic strains of Actinobacillus actinomycetemcomitans. J Periodontal Res 1981; 16: 379-389.
3. Listgarten M, Lai C, Evian, I. Comparative antibody titers to Actinobacillus actinomycetemcomitans in juvenile periodontitis, chronic periodontitis and periodontally health subjects. J Clin Periodontol 1981; 8: 155-164.
4. Nowotny A, Behling U, Hammond B, Lai C, Listgarten M, Pham P, Sanavi F. Release of toxic microvesicles by Actinobacillus actinomycetemcomitans. Infect Immun 1982; 37: 151-154.
5. Tsai C, Shenker B, DiRienzo J, Malmud D, Taichman N. Extraction and isolation of a leukotoxin from Actinobacillus actinomycetemcomitans with polymyxin B. Infect Immun 1984; 43: 700-705.
6. Zambon JJ. Actinobacillus actinomycetemcomitans in human periodontal disease. J Clin Periodontol 1985; 12: 1-20.
7. Moore W. Microbiology of periodontal disease. J Periodontal Res 1987; 22: 335-341.
8. Slots J, Listgarten M. Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. J Clin Periodontol 1988; 15: 85-93.
9. Preus H, Zambon J, Dunford R, Genco, R. The distribution and transmission of A. actinomycetemcomitans in families with established adult periodontitis. J Periodontol 1994; 65: 2-7.
10. Alaluusua S, Asikainen S. Detection and distribution of Actinobacillus actinomycetemcomitans in the primary dentition. J Periodontol 1988; 59: 504-507.
11. Asikainen S, Lai C, Alaluusua S, Slots S. Distribution of Actinobacillus actinomycetemcomitans serotypes in periodontal health and disease. Oral Microbiol Immunol 1991; 6: 115-118.
12. Mandell R, Socransky S. A selective medium for isolation Actinobacillus actinomycetemcomitans and the incidence of the organism in juvenile periodontosis. J Periodontol 1981; 52: 593-598.
13. Slots J, Reynolds H, Genco R. Actinobacillus actinomycetemcomitans in human periodontal disease. A cross-sectional microbiological examination. Infect Immun 1980; 29: 1013-1020.
14. Ashley F, Gallagher J, Wilson R. The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, Bacteroides intermedius and spirochaetes in the subgingival microflora of adolescents and their relationship with the amount of supragingival plaque and gingivitis. Oral Microbiol Immunol 1988; 3: 77-82.
15. Asikainen S, Alaluusua S, Kari K, Kleemola-Kujala E. Subgingival microflora and periodontal conditions in healthy teenagers. J Periodontol 1986; 57: 505-509.
16. Delaney JE, Kornman KS. Microbiology of subgingival plaque from children with localized prepuberetal periodontitis. Oral Microbiol Immunol 1987; 2: 71-76.
17. Zambon J, Slots J, Genco R. Serology of oral Actinobacillus actinomycetemcomitans and serotype distribution in human periodontal disease. Infect Immun 1983; 41: 19-27.
18. Saarela M, Asikainen S, Alaluusua S, Asikainen T, Pyhala L, Lai C, Jousimies-Somer H. Frequency and stability of mono- or poly-infection by Actinobacillus actinomycetemcomitans serotypes a,b,c,d or e. Oral Microbiol Immunol 1992; 7: 277-279.
19. Mombelli A, Gmur R, Lang N, Corbet E, Frey J. Actinobacillus actinomycetemcomitans in Chinese adults. Serotype distribution and analysis of the leukotoxin gene promoter locus. J Clin Periodontol 1999; 26: 505-510.
20. Doğan B, Saarela M, Jousimies-Somer H, Alaluusua S, Asikainen S. Actinobacillus actinomycetemcomitans serotype e - biotypes, genetic diversity and distribution in relation to periodontal status. Oral Microbiol Immunol 1999; 14: 98-103.
21. Laiko L, Kuula H, Doğan B, Asikainen S. Actinobacillus actinomycetemcomitans proportion of subgingival bacterial flora in relation to its clonal type. Eur J Oral Sci 2002; 110: 212-217.
22. Gmür R, McNabb H, van Steenbergen TJM, Baehni P, Mombelli A, Van Winkelhoff A, Guggenheim B. Seroclassification of hitherto nontypeable Actinobacillus actinomycetemcomitans strains: evidence for a new serotype e. Oral Microbiol Immunol 1993; 8: 116-120.
23. Zambon J, Umemoto T, De Nardin E, Nakazawa F, Christersson L, Genco R. Actinobacillus actinomycetemcomitans in the pathogenesis of human periodontal disease. Adv Dent Res 1988; 2: 269-274.
24. Chung H, Chung C, Son S, Nisengard R. Actinobacillus actinomycetemcomitans serotypes and leukotoxicity in Korean localized juvenile periodontitis. J Periodontol 1989; 60: 506-511.
25. Gmür R, Baehni P. Serum immunoglobulin G responses to various Actinobacillus actinomycetemcomitans serotypes in a young ethnographically heterogenous periodontitis patient group. Oral Microbiol Immunol 1997; 12: 1-10.
26. Asikainen S, Chen C, Slots J. Actinobacillus actinomycetemcomitans genotypes in relation to serotypes and periodontal status. Oral Microbiol Immunol 1995; 10: 65-68.
27. Zambon J, Haraszthy V, Hariharan G, Lally E, Demuth D. The microbiology of early-onset periodontitis: Association of highly toxic Actinobacillus actinomycetemcomitans strains with localized juvenile periodontitis. J Periodontol 1996; 67: 282-290.
28. Tinoco EMB, Stevens R, Haubek D, Lai C-H, Balachandran S, Preus H. Relationship of serotype, leukotoxin gene type and lysogeny in Actinobacillus actinomycetemcomitans to periodontal disease status. Eur J Oral Sci 1997; 105: 310-317.
29. Goldsworthy N. Bacterium actinomycetemcomitans: a description of two strains. J Pathol Bacteriol 1938; 46: 207-208.
30. Holm P. Studies on the etiology of human Actinomycosis. I. The other microbes of Actinomycosis and their importance. Acta Pathol Microbiol Scand 1950; 27: 736-751.
31. Holm P. Studies on the etiology of human Actinomycosis. II. Do the other microbes of Actinomycosis possess virulence? Acta Pathol Microbiool Scand 1951; 28: 391-406.
32. Vallee A, Gailland J. Infection pyogene catagiense de`la souris determince par Bacillus actinomycetemcomitans. Ann Inst Pasteur Lille 1953; 84: 647-649.
33. Kaplan AH, Weber DJ, Odone EZ, Perfect JR. Infection due to Actinobacillus actinomycetemcomitans: 15 cases and review. Rev Infect Dis1989; 11: 46-63.
34. Kristinsson KG, Thoreirsson G, Holbrook WP. Actinobacillus actinomycetemcomitans and endocarditis. J Infect Dis 1988; 157: 599.
35. Verhaaren H, Claeys G, Verschraegen G, de Niel C, Leroy J, Clement D. Endocarditis from a dental focus. Importance of oral hygiene in valvar heart disease. Int J Cardiol 1989; 23: 343-347.
36. Burgher L, Loomis G, Ware F. Systemic infections due to Actinobacillus actinomycetemcomitans. Am J Clin Pathol 1973; 60: 412-415.

37. Townsend T, Gillenwater J. Urinary tract infection due to Actinobacillus. J Am Med Assoc 1969; 210: 558.
38. van Winkelhoff AJ, Overbeek BP, Pavicic MJAMP, van der Bergh PA, Ernst JPMG, de Graaff J. Long-standing bacteremia caused by oral Actinobacillus actinomycetemcomitans in a patient with a pacemaker. Clin Infect Dis 1993; 16: 216-218.
39. Yuan A, Yang PC, Lee LN, Chang DB, Kuo SH, Luh KT. Actinobacillus actinomycetemcomitans pneumonia with chest wall involvement and rib destruction. 1992; 101: 1450-1452.
40. Haubek D, Poulsen K, Asikainen A, Kilian M. Evidence for absence in northern Europe of especially virulent clonal types of Actinobacillus actinomycetemcomitans. J Clin Microbiol 1995; 33: 395-401.
41. Poulsen K, Theilade E, Lally E, Demuth D, Kilian M. Population structure of Actinobacillus actinomycetemcomitans: a framework for studies of disease-associated properties. Microbiol 1994; 140: 2049-2060.
42. Slots S, Ting M. Actinobacillus actinomycetemcomitans and Porpyromonas gingivalis in human periodontal disease: occurrence and treatment. Periodontol 2000 1999; 20: 82-121.
43. Socransky S, Haffajee AD, Ximenez-Fyvie LA, Feres M, Mager D. Ecological considerations in the treatment of Actinobacillus actinomycetemcomitans and Porpyromonas gingivalis periodontal infections. Periodontol 2000 1999; 20: 341-362.
44. Caton J. Periodontal diagnosis and diagnostic aids. In: Proceedings of the World Workshop in Clinical Periodontics. Chicago: The American Academy of Periodontology; 1989; I/1-I/22.
45. The American Academy of Periodontology. Consensus report and summary of plenary session. Periodontal diagnosis and diagnostic aids. In: Proceedings of the World Workshop in Clinical Periodontics. Chicago: The American Academy of Periodontology; 1989; I/23-I/32.
46. Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999; 4:1-6.
47. Möller A. Microbiological examination of root canals and periapical tissues of human teeth. Odontol Tidskr 1966; 74: 1-380.
48. Slots J. Selective medium for isolation of Actinobacillus actinomycetemcomitans. J Clin Microbiol 1982; 15: 606-609.
49. Lai C, Listgarten M, Shirakawa M, Slots J. Bacteroides forsythus in adult gingivitis and periodontitis. Oral Microbiol Immunol 1987; 2: 152-157.
50. Lai C, Oshima K, Slots J, Listgarten M. Wolinella recta in adult gingivitis and periodontitis. J Periodontal Res 1992; 27: 8-14.
51. Lai C-H, Listgarten M. Immune labeling of certain strains of Actinomyces naeslundii and Actinomyces viscosus by fluorescence and electron microscopy. Infect Immun 1979; 25: 1016-1028.
52. Lai C-H, Listgarten MA, Hammond BF. Comparative ultrastrcture of leukotoxic and non-leukotoxic strains of Actinobacillus actinomycetemcomitans. J Perodontal Res 1981; 16: 379-389.
53. Baehni P, Tsai C-C, McArthur WP, Hammond BF, Taichman NS. Interaction of inflammatory cell and oral microorganisms. VIII. Detection of leukotoxic activity of a plaque-derived gram-negative microorganism. Infect Immun 1979; 24: 233-243.
54. Taichman NS, Iwase M, Korchak HM, Berthold P, Lally E. Membranolytic of Actinobacillus actinomycetemcomitans leukotoxin. J Perodontal Res 1991; 26: 258-260.
55. Lally ET, Golub EE, Kieba IR, Taichman NS, Decker S, Berthold P, Gibson CW, Demuth DR, Rosenbloom J. Structure and function of the B and genes of Actinobacillus actinomycetemcomitans leukotoxin complex. Microbial Pathogenesis 1991; 11: 111-121.
56. Yoshida Y, Suzuki N, Nakano Y, Shibuya K, Ogawa Y, Koga T. Distribution of Actinobacillus actinomycetemcomitans serotypes and Porphyromonas gingivalis in Japanese adults. Oral Microl Immunol 2003; 18: 135-139.
57. Yamamoto M, Nishihara T, Koseki T, He T, Yamato K, Zhang Y, Nakahima K, Oda S, Ishikawa I. Prevalence of Actinobacillus actinomycetemcomitans serotypes in Japanese patients with periodontitis. J Periodontal Res 1997; 32: 676-681.
58. Paju S, Saarela M, Alaluusua S, Fives-Taylor P, Asikainen S. Characterization of serologically nontypeable Actinobacillus actinomycetemcomitans isolates. J Clin Microbiol 1998; 36: 2019-2022.
59. Kaplan JB, Perry MB, MacLean LL, Furgang D, Wilson ME, Fine DH. Structural and genetic analyses of O polysaccharide from Actinobacillus actinomycetemcomitans serotype f. Infect Immun 2001; 69: 5375-5384.
60. Slots J, Rams T Microbiology of periodontal disease. In: Slots J, Taubman M, ed. Contemporary oral microbiology and immunology. St Louis, MS: Mosby, 1992; 425-443.
61. Slots J. Bacterial specificity in adult periodontitis. J Clin Periodontol 1986; 13: 912-917.
62. Haffajee AD, Cugini MA, Tanner A, Pollack RP, Smith C, Kent RL, Socransky SS. Subgingival microbiota in healthy, well-maintained elder and periodontitis subjects. J Clin Periodontol 1998; 25: 346-53.
63. Wolff LF, Aeppli DM, Pihlstrom B, Anderson L, Stoltenberg J, Osborn J, Hardie N, Shelburne C, Fischer G. Natural distribution of 5 bacteria associated with periodontal disease. J Clin Periodontol 1993; 20: 699-706.
64. Hamlet AM, Cullinan MP, Westerman B, Lindeman M, Bird PS, Palmer J, Seymour GJ. Distribution of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in an Australian population. J CliniPeriodontol 2001; 28: 1163-1171.
65. Furcht C, Eschrich K, Merte K. Detection of Eikenella corrodens and Actinobacillus actinomycetemcomitans by use of the polymerase chain reaction (PCR) in vitro and in subgingival plaque. J Clin Periodontol 1996; 23: 891-897.
66. Umeda M, Chen C, Bakker I, Contreras A, Morrison JL, Slots J. Risk indicators for harboring periodontal pathogens. J Periodontol 1998; 69: 1112-1119.