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標題: | 犬貓螺旋樣桿菌的研究 Study of Gastric Helicobacter-Like Organisms (GHLOs) in Dogs and Cats |
作者: | Ping-Hsu Chen 陳炳旭 |
指導教授: | 萬灼華(Cho-Hua Wan) |
共同指導教授: | 黃慧璧(Hui-Pi Huang) |
關鍵字: | 胃螺旋樣桿菌,犬貓胃螺旋樣桿菌盛行率,人畜共傳的胃螺旋樣桿菌,臺灣犬貓的胃螺旋樣桿菌, canine helicobacteriosis,feline helicobacteriosis,Helicobacter generic PCR assay,GHLOs prevalence,zoonotic helicobacteriosis, |
出版年 : | 2009 |
學位: | 碩士 |
摘要: | 螺旋桿菌 (Helicobacter)為微好氧性的革蘭氏陰性菌,外型為彎曲至螺旋狀,大小約0.5×5-10 μm,具有運動性。在1983年,Marshall 與Warren經實驗感染,成功從自已的胃生檢樣本中培養出Helicobacter pylori,並證實此細菌會引起胃潰瘍疾病。後續的研究發現,H. pylori的存在會引起急性或慢性胃炎,並與胃潰瘍和十二指腸潰瘍、胃腺癌 (gastric adenocarcinoma)及胃黏膜相關淋巴組織淋巴瘤(mucosa-associated lymphoid tissue lymphoma;MALT lymphoma)相關。自H. pylori的發現後,非H. pylori的胃螺旋樣桿菌 (gastric helicobacter-like organisms, GHLOs)陸續在許多不同動物種類的胃中被發現。至目前為止,至少已有38種的不同的螺旋桿菌被分類出來,但還有很多仍在鑑定中。螺旋桿菌又依據細菌生長位置,可細分為二大族群: 胃螺旋桿菌群 (gastric helicobacters) 與腸肝螺旋桿菌群 (enterohepatic helicobacters)。在犬貓的GHLOs感染中,通常多無臨床症狀,但於嚴重感染時,可發現慢性活化性胃炎 (chronic active gastritis)。在近代的研究文獻中,發現犬貓感染GHLOs的盛行率極高 (41%-100%),且其中有數種non-H. pylori GHLOs亦曾由人與動物胃中被分離鑑定,升高人跟寵物間有人畜共傳GHLOs的可能性。目前在台灣,犬貓的non-H. pylori GHLOs感染盛行率、感染細菌種別及人畜共傳螺旋桿菌研究的資料仍非常有限。本次實驗主要目的是利用分子生物檢測診斷方法,以瞭解台灣犬貓GHLOs的感染的狀況及盛行率,並鑑定目前在台灣犬貓GHLOs種別,評估是否有人畜共傳的危機存在,更希望可同時建立臺大動物醫院犬貓GHLOs例行性的診斷鑑定方法,以提供臨床醫師在臨床的診斷依據及未來在犬貓GHLOs人畜共傳疾病研究的參考。本實驗乃收集臺灣大學生農學院附設動物醫院36個犬貓胃生檢樣本及剖解病材,利用分子生物學診斷方法,於18個胃檢體中偵測到GHLOs (陽性率為50%)。將其中17個陽性樣本,進行16S rRNA基因增幅與定序,經序列比對,發現17個GHLOs其基因序列均與胃螺旋桿菌相似性高,並歸納為GHLO。於進一步的基因序列比較分析中,這17個GHLOs與“H. heilmannii”、H. bizzozeronii、H. felis、H. salomonis、H. cynogastricus及H. baculiformis較為相似 (相同度為97%-99%),但與H. pylori及肝腸螺旋桿菌群細菌差異較大,其相同度分別為93.9%-94.5%及91.7%-93.8%。在親緣關係分析結果顯示,其中13個GHLOs與“H. heilmannii”、H. bizzozeronii、H. felis及H. salomonis最為相似,由於“H. heilmannii”、H. bizzozeronii、H. felis及H. salomonis曾在人及動物胃中被分離檢出過,因此我們懷疑這13個GHLOs亦可能存有人畜共傳的危險。而另外4個GHLOs與目前已知的Helicobacter差異較大,應是尚未檢出命名的Helicobacters或是已知菌種的臺灣分株,此4個GHLOs是否存在人類及其他動物胃中,或是否具有人畜共傳的可能性,目前仍不清楚,需再進一步研究釐清。於此研究中利用分子診斷技術,可快速準確偵測犬貓胃中GHLOs的存在,經由序列比對,可大致鑑別GHLOs的種別,可快速準確提供臨床醫師在臨床的診斷依據,評估人畜共傳的危險性,並可減少犬貓GHLOs人畜共傳的機會。 Helicobacteriosis is one of the most prevalent infectious diseases in humans and animals. Multiple gastric helicobacter-like organisms (GHLOs) have been found in human and many animal species and some of them have been reported to cause diseases in humans, dogs, and cats. In Taiwan, the prevalence of helicobacter infection in dogs and cats is still unclear and the zoonotic research of helicobacteriosis is also very limited. The purpose of this study is using PCR assay to identify the helicobacter infected dogs and cats in Taiwan, to determine the prevalence of helicobacteriosis in pet dogs and cats, and to evaluate the zoonotic risk of helicobacter infection between pets and humans. In this study, tissues collected via gastric biopsy or at necropsy from 36 pets were screened by the Helicobacter generic PCR assay and positive samples were further analyed for the sequence of 16S rRNA gene. A total of 18 samples were found to be helicobacter positive on helicoabcters based on the Helicobacter generic PCR result (prevalence of 50%). The 16S rRNA gene sequences of 17 samples were determined and compared to the of previously reported gastric helicobacters. The analysis results revealed the 13 samples were closely related to previously identified zoonotic helicobactes, 6 samples similar to “H. heilmannii”, 5 samples similar to “H. heilmannii” and H. bizzozeronii and 2 samples similar to H. felis and H. salomonis. Four different samples were distinctly different from currently known gastric helicobacters in dogs and cats and were suggested to be newly identified GHLOs. In conclusion, gastric helicobacteriosis is highly prevalent in pet dogs and cats in Taiwan. Potential risks of zoonotic helicobacteriosis of dogs, cats, and humans are at present uncertain, but with existing possibility. Several newly identified GHLOs are recognized and further study is required for the pathogenesis and zoonotic risk of these GHLOs. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43808 |
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