- 學位論文
利用巣式聚合酶連鎖反應調查台北市犬艾利希氏體病
Survey on canine ehrlichiosis in Taipei by nested PCR
摘要
犬艾利希氏體病，是一種由立克次體所引起的疾病。由於感染時的臨床症狀不具特異性，往往不易診斷。臨床上，目前對於診斷犬艾利希氏體病主要仍然依靠光學顯微鏡鏡檢及商品化kit為主，但是都有其缺點。本研究選擇以巢式聚合酶連鎖反應(nested-PCR)的方式，針對不同種類犬艾利希氏體之16S rRNA基因序列作引子設計，以調查台北市各動物醫院所送檢之疑似病例感染情況，並對臨床上造成的病害及和焦蟲混合感染的情況作了解。目前已由所收集的174個疑似病例中檢驗出29個E. canis及2個E. platys之陽性病例，並定序確認。此外亦收集了台北市動物之家所提供的流浪犬隻血液作檢驗，初步於31個血液樣本中，分別檢測出4，11及2例E. canis，E. platys及E. equi陽性病例。在16S rRNA基因序列分析上，也已定序出台北地區E. canis及E. platys之全長約1400bp的16S rRNA基因序列，並以DNASTAR 程式(DNASTAR Inc., Madison, WI)和國外已發表之序列作親緣性分析。
關鍵字
艾利希氏體並列摘要
Canine ehrlichiosis is a tick-borne disease caused by obligate intracellular parasites of the genus Ehrlichia. The diagnosis is difficult, because the clinical signs of canine ehrlichiosis are always not specific. Microscope and commercial kit are usually used for clinical diagnosis. However, some disadvantages exist in these diagnostic methods. We established nested-PCR with primers derived from the 16S rRNA gene sequence to study the infection rate, pathology, and coinfection with Babesia spp. of suspicious sample from animal hospitals in Taipei and Taipei Municipal Animal Shelter. We had designed five Ehrlichia-species-specific primers, and were able to amplify E. canis (29/174)and E. platys(2/174) of owned dogs as well as E. canis (4/31), E. platys(11/31)and E. equi (2/31) of stray dogs sampled from Taipei Municipal Animal Shelter. At the same time, we have sequenced the full 16S rRNA gene sequences of E. canis and E. platys, and compared with the other strains in gene bank by DNASTAR.