Differentiation of Four Eimeria Species Commonly Seen in Taiwanese Rabbits by RAPD

臺灣常見兔艾美球蟲的RAPD分子鑑定

李明憲(Ming-Hsien Li) ； 廖朝財(Chao-Tsai Liao) ； 歐威志(Wei-Chih Ou) ； 陳碧蓮(Pei-Lain Chen) ； 黃瓊華(Chiung-Hua Huang) ； 陳妤瑄(Yu-Hsuan Chen) ； 黃海義(Hai-I Huang) ； 黃鴻堅(Hong-Kean Ooi)

《臺灣獸醫學雜誌》 38卷4期 (2012/12) Pp. 227-232

https://doi.org/10.7009/TVJ.201212.0227

摘要

以11個非特異性引子（G2，E111-E120）進行隨機增幅多型性DNA（RAPD）鑑別4 種兔艾美球蟲（Eimeria piriformis、E. magna、E. perforans及E. media）。以次氯酸鈉處理及溶解緩衝液作用，再以玻璃珠將已芽孢化卵囊均質化後萃取DNA。11個引子中，E113、E114及E120對E. piriformis皆可產生獨特的電泳帶，此可用於與E. magna、E. media、E. perforans區別。而E111則在E. magna、E. media及E. perforans可產生不同的電泳帶，在E. piriformis則無。因此，合併E111與E113、E114及E120的其中一個引子便可將4種艾美球蟲加以鑑別。可知，隨機增幅多型性DNA技術可用於輔助傳統型態學鑑別方法之不足。

關鍵字

隨機增幅多型性DNA ； 艾美球蟲 ； 鑑定 ； 兔

並列摘要

To differentiate four Eimeria species from rabbit, namely, Eimeria piriformis, E. magna, E. perforans, and E. media, random amplified polymorphic DNA (RAPD) with 11 sets of non-specific primers, designated G2, E111 to E120, was carried out. Modification of DNA extraction from sporulated oocyst using sodium hypochlorite treatment and lysis buffer followed by glass beads homogenization was found to yield a better result. Of the 11 primer sets tested, 3 (E113, E114, and E120) were able to produce unique band patterns for E. piriformis, which can be used to differentiate it from E. magna, E. media, and E. perforans. However, primer E111 was able to produce different band patterns for E. magna, E. media, and E. perforans but no band for E. piriformis. Thus, using a combination of E111 with either E113 or E120 primers, we can differentiate each of the 4 species from one another. Thus, RAPD has the potential to be used as a molecular method for confirming and differentiating the identity of the Eimeria species from rabbit to complement the classical morphometric method.

並列關鍵字

RAPD ； Eimeria ； Differentiation ； rabbit