台灣國人牙周病原菌之分析

Analysis of Periodontal Pathogens in Taiwan

林姸華(Yen-Hua Lin)

指導教授 ： 賴辰雄 ； 何坤炎

高雄醫學大學/口腔醫學院/牙醫學研究所/碩士(2008年)

https://doi.org/10.6832/KMU.2008.00124

摘要

背景:
不同地區的牙菌斑組成不盡相同，目前的資料尚無足夠的台灣代表性，需要收集更多菌種。本研究將倂用兩種檢測方式，分析多種致病菌與慢性牙周炎之關係。

研究目的:
1.比較Porphyromonas gingivalis (P. gingivalis)、Tannerella forsythia (T. forsythia; previously Tannerella forsythensis或Bacteroides forsythus)、Treponema denticola (T.denticola)、Eikenella corrodens (E. corrodens)、Campylobacter rectus(C. rectus; previously Wollinella recta)、Prevotella intermedia (P. intermedia)、Prevotella nigrescens (P. nigrescens)在輕微牙齦炎與牙周炎牙齦上和牙齦下牙菌斑之組成百分比及盛行率。
2.探討組成百分比與牙周探測深度(Probing depth, PD)、臨床附連喪失(Clinical attachment loss, CAL)、牙菌斑指數(Plaque Index, PI)、牙齦指數(Gingival Index, GI)之相關性。
3.分析間接免疫螢光試驗(indirect immunofluorescence assay,IF)和聚合酶連鎖反應(polymerase chain reaction, PCR)對上述菌種之敏感度及特異性。

材料與方法:
以51位中至重度慢性牙周炎患者為研究對象，依其疾患狀況採集下列3種部位:輕微牙齦炎的牙齦上、牙周炎的牙齦上與牙齦下牙菌斑。以IF計算菌種之組成比例及盛行率，並用PCR確認。使用ANCOVA、Chi-square (卡方檢定)、Pearson correlation coefficient及simple linear regression (簡單線性迴歸)作統計分析。

結果:
各菌種在不同部位的組成百分比均有顯著差異(p<0.05)，P.gingivalis、T. forsythia、T. denticola、C. rectus在牙周炎牙齦下部位比例最高(8.50、3.00、3.82、0.84%)，P. intermedia、P. nigrescens以牙周炎牙齦上部位最高(3.22、4.18%)，E.corrodens則以牙齦炎部位最高(1.12%)。各菌種的IF盛行率皆以牙周炎牙齦下部位最高，E. corrodens、C. rectus分別為92.16和98.04%，其餘均是100.00%。不同部位均有顯著差異(p<0.05)。PCR和IF的一致性百分比除了T. forsythia、P. nigrescens之外，皆在73.57~83.57%。以PCR為參考時，IF的敏感度為77.34~100.00%，特異性為0.00~33.33%。以IF為參考時，PCR的敏感度除了T. forsythia和P. nigrescens之外，皆大於81.45%，特異性除了T. forsythia和P. intermedia為100%之外，其餘皆在50.00%以下。P. gingivalis、T. forsythia、T. denticola、 P.intermedia、P. nigrescens、C. rectus的組成比例皆與PD、CAL、PI及GI呈正相關。唯有E.corrodens和PD、CAL及GI呈負相關，其中與PD、CAL有達顯著差異(p<0.05)。將E. corrodens的組成比例和年齡作簡單線性迴歸分析，顯示隨著年齡增加，E. corrodens的百分比會降低。

結論:
P. gingivalis、T. forsythia、T. denticola、C. rectus、P. intermedia、P. nigrescens與台灣慢性牙周炎較有關聯。值得注意的是P. intermedia和P. nigrescens的組成比例在牙周炎牙齦上比牙齦下還高。平均年齡47.63~48.45歲的慢性牙周炎中，P.gingivalis、T. forsythia、T. denticola、C. rectus、P. intermedia、P. nigrescens的組成比例與齒槽骨破壞及牙齦發炎呈正相關。E. corrodens依多數研究指出和年輕型牙周炎有關，本研究進一步顯示E. corrodens和成人牙周炎相關性較低。雖然大部分的菌種在IF和PCR的一致性都有接近80%，但是T. forsythia和P.nigrescens偏低，因此仍以多種檢測來確認較為可靠。

關鍵字

慢性牙周炎 ； 牙菌斑 ； 間接免疫螢光試驗 ； 聚合酶連鎖反應

並列摘要

Backgrounds:
Composition of microbiota in dental plaque may differ from geographical regions. Currently, there is little information available regarding periodontal pathogen profile for Taiwan individuals. This study focused on the investigation of the composition of periodontal pathogens in Taiwan subjects with chronic periodontitis.

Objectives:
1. To compare the proportions and prevalences of both supragingival and subgingival plaque in chronic periodontitis site and plaque samples from mild gingivitis. Periodontal pathogens for this study were targeted on Porphyromonas gingivalis(P. gingivalis), Tannerella forsythia (T. forsythia; previously Tannerella forsythensis or Bacteroides forsythus), Treponema denticola(T.denticola),Eikenella corrodens (E. corrodens), Campylobacter rectus (C. rectus; previously Wollinella recta), Prevotella intermedia(P. intermedia),Prevotella nigrescens (P. nigrescens).
2. To determine the correlation coefficient between the proportion of each tested periodontal pathogen and clinical parameters including probing depth (PD), clinical attachment loss (CAL), plaque index (PI) and gingival index (GI).
3.To define the sensitivity and specificity of indirect immunofluorescence assay (IF) and polymerase chain reaction (PCR) for detection of the targeted periodontal pathogens.

Material and Methods:
A total of 51 subjects with moderate to severe chronic periodontitis were recruited. Supra and/or subgingival plaque samples were collected from sites with chronic periodontitis and mild gingivitis. The targeted pathogens were detected by both IF and PCR. The statistical analysis was performed by ANCOVA, Chi-square, Pearson correlation coefficient and simple linear regression test.

Results:
Proportions of targeted species were significantly different among sites (p<0.05). Subgingival proportions of P. gingivalis, T. forsythia,T. denticola, C. rectus were highest as compared to supragingival of chronic periodontitis and mild gingivitis sites (8.50, 3.00, 3.82 and 0.84% respectively). In contrast to above pathogens, elevated proportions of P. intermedia and P. nigrescens were noted in supragingival periodontitis area as compared to subgingival (3.22 and 4.18% respectively).Proportion of E. corrodens was higher in gingivitis site as compared to
periodontitis both in supra and subgingival site (1.12%). Based on IF results, frequency of E. corrodens and C.rectus were 92.16 and 98.04% respectively. All other targeted species occurred 100% in subgingival periodontitis site. With the exception of T. forsythia and P. nigrescens, the agreement between both PCR and IF was about 73.57~83.57%. When PCR was used as the reference, the sensitivity of IF was 77.34~100.00%. Specificity was 33.33%. On the other hand, when IF was used as the reference, the sensitivity of PCR was≥81.45%,except for T. forsythia and P.nigrescens. The specificity of most tested periodontal pathogens was ≤50.00%. However, T. forsythia and P. intermedia had 100% specificity.The proportions of P. gingivalis, T. forsythia, T. denticola,P. intermedia, P. nigrescens, C. rectus were all in positive correlation with PD, CAL, PI and GI.However, E. corrodens had negative correlation with PD, CAL and GI.The proportion of E. corrodens and age was analyzed by simple linear regression. The result showed that the proportion of E. corrodens was decreased when the age increased.

Conclusions:
The present study demonstrated that P. gingivalis, T.forsythia,T. denticola, C. rectus, P. intermedia, P.nigrescens were the major periodontal pathogens for adult chronic periodontitis in Taiwan.It was worthwhile to note that the proportions of P.intermedia and P.nigrescens were higher in supragingival than subgingival periodontitis site.The proportion of P.gingivalis, T.forsythia, T.denticola, C. rectus,P.intermedia, P.nigrescens were in positive correlation with alveolar bone loss and gingival inflammation in chronic periodontitis.According to other literatures, E. corrodens had positive relationship with juvenile periodontitis. Our study further indicated that it had less correlation with adult periodontitis. Although most other periodontal pathogens had nearly 80% of agreement between IF and PCR, it was lower in T. forsythia and P. nigrescens. This study suggests that it is wisely to use multiple detection methods for clinical diagnosis of specific periodontal pathogens.

並列關鍵字

Chronic periodontitis ； Dental plaque ； Indirect immunofluorescence assay ； Polymerase chain reaction

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