運用巢式聚合酵素鍊反應偵測犬吉貝氏焦蟲

外文標題: 
Detection of Babesia gibsoni by nest polymerase chain reaction
校院系所: 
國立臺灣大學 獸醫學研究所
指導教授: 
費昌勇
出版年份: 
1999年
主題類別: 
摘要: 

本實驗研究主要目的在於使用PCR的技術區分犬吉貝氏焦蟲與型態學相類似的犬血巴東蟲或賀-邱氏小體。目前對於診斷犬吉貝氏焦蟲主要仍然依靠光學顯微鏡鏡檢為主,但是這個方法的敏感性太低,應用分子生物技術可以提供較精確的診斷。本實驗研究巢式聚合酵素鏈反應與DNA探針墨點雜合法,以發展新的方法做為犬吉貝氏焦蟲診斷。利用16S rDNA序列設計了兩組引子以進行巢式聚合酵素鏈反應,巢式聚合酵素鏈反應可以偵測到DNA量之敏感度比DNA探針高了1000倍。我們藉由所蒐集的14 個吉貝氏焦蟲樣本進行定序分析,瞭解到台灣地區犬吉貝氏焦蟲的16S rDNA部分序列,與基因庫上發表的序列相似性只有73%,顯示出台灣地區犬吉貝氏焦蟲的序列與國外的序列有著顯著的差異。

外文摘要: 

The objective of this study was to determine whether different isolates of Babesia gibsoni could be distinguished from morphologically similar isolates of Haemobartonella canis or Howell-Jolly body by using the polymerase chain reaction technology. At present the diagnosis of Babesia gibsoni is by microscope. This sensitivity of this method is too low. There is need for improved diagnostic aids to facilitate recogniziation and treatment of this parasite infection. This study which apply molecurar biological technology ─nest polymerase chain reaction(nest PCR) and DNA probe Dot blotting hybridization that detect the canine Babesia gibsoni successfully. Two sets of primer designed from the 16S ribosomal DNA gene sequences(Gene bank B.G) were used to amplify the genomic DNA of Babesia gibsoni by nest PCR. The sensitivity of nest PCR is 1000-fold higher than DNA Dot blotting hybridization. Hence, the nest PCR assay supersedes the DNA probe and older detection methods for the detection of Babesia gibsoni in the blood sample. We collected 14 samples of Babesia gibsoni DNA to sequencing. Analysis for the partial sequence of 16S rDNA of the Babesia gibsoni in Taiwan(Taiwan B.G). The similarity between the Taiwan B.G and the Gene bank B.G are 73%. There is major difference between the sequence of the Babesia gibsoni from those of the Taiwan and other countries.