牙周炎為世界上最盛行的口腔疾病之一，主要由牙菌斑細菌所導致，其中，牙齦卟啉單胞菌、齒垢密螺旋體及福賽斯坦納菌被認定為牙周炎致病細菌。此外，口腔原蟲：齒齦阿米巴及口腔鞭毛蟲較常出現於口腔衛生不佳或牙周炎患者口腔內，然而目前的研究無法證實兩者和牙周炎間的關聯，同時台灣也並未有口腔原蟲的相關報導研究，因此，本研究旨在調查口腔原蟲在台灣牙周炎患者之盛行率，並探討口腔原蟲和口腔致病細菌間的作用關係之於牙周炎的影響。我們自30位牙周炎患者口腔中，分別採集輕微牙齦炎及嚴重牙周炎部位之牙菌斑檢體，並以聚合酶連鎖反應偵測口腔原蟲及口腔致病細菌。在60個檢體中，齒齦阿米巴感染率為73.33%，亞型比例分別為13.33%的ST1、13.33%的ST2-kamaktli及46.67%同時感染兩種亞型，口腔鞭毛蟲感染率為23.33%，齒齦阿米巴及口腔鞭毛蟲之共同感染率則為18.33%。而根據統計分析結果，口腔鞭毛蟲和福賽斯坦納菌的感染率在感染症狀較為嚴重的部位明顯高於輕微部位 (p < 0.05) 。在三種口腔致病細菌中，牙齦卟啉單胞菌感染率最高，高達83.33%，然而與口腔鞭毛蟲之共同感染率卻為三者最低。為了釐清兩者間的關係，我們利用共同培養實驗測試牙齦卟啉單胞菌與口腔鞭毛蟲之生長情況，結果顯示口腔鞭毛蟲數量隨著時間有顯著下降之趨勢。我們也發現以口腔鞭毛蟲之條件培養基培養48小時的牙齦卟啉單胞菌致病基因 (hagB) 表現量顯著上升。另一方面，在細胞損傷能力測試中，口腔鞭毛蟲相對於控制組有顯著的細胞破壞能力，而牙齦卟啉單胞菌之條件培養基及裂解液亦同樣具有破壞細胞之能力。作為台灣第一篇口腔原蟲之研究，我們調查了口腔原蟲於台灣牙周炎患者之盛行率，並發現口腔原蟲與口腔致病細菌間之於牙周炎的潛在作用關係。

To investigate the prevalence and the correlates of the oral microbes among patients with chronic periodontitis in Taiwan, we collected 60 clinical samples from 30 patients in NCKU hospital to detect the three oral bacteria and two protozoans by polymerase chain reaction (PCR). In this population, E. gingivalis was present in 44 samples (73.33%), while T. tenax was present in 15 samples (23.33%). Besides, the prevalence of P. gingivalis was 83.33%, T. denticola 78.33%, and that of T. forsythia was 80.0%. While collecting dental plaques from sites with mild gingivitis and chronic periodontitis from each patient, we found T. tenax and T. forsythia infection were significantly higher in chronic positions than in mild positions. Moreover, from the analysis of these results, we noticed that the infection rate of T. tenax and P. gingivalis co-infections (18.33%) was the lowest compared to other two bacteria even though the prevalence of P. gingivalis was the highest. However, in our in vitro studies, there was no significant change in the expression level of the virulence factors of P. gingivalis while T. tenax co-culturing with P. gingivalis. We also showed pathogenesis of T. tenax by cytopathic effect test. In conclusion, we found the prevalence of two oral protozoans in Taiwan and their potential interaction with pathogenic bacteria on periodontitis. As the first study to detect E. gingivalis and T. tenax among patients with periodontitis in Taiwan, it revealed a new way for research on periodontitis and oral parasites.

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