我国犬埃立克体病病原的分离与鉴定 Ⅰ.病原16S rRNA基因序列测定与分析

Isolation and Identification of the Causative Agent of Canine Ehrlichiosis Found in China Ⅰ.Sequencing and Analyzing of 16S rRNA Gene

用实验动物接种法，分离保存我国某养犬基地犬埃立克体病病原。以此病原对易感犬接毒，接毒犬发病后采集急性期全血提取基因组，对病原 １６ Ｓｒ Ｒ Ｎ Ａ 基因进行扩增、克隆测序。结果，病原被保存在病犬血细胞内，并能成功地使健康犬感染发病；检测到的 ２ 种致犬埃立克体病病原 １６ Ｓ ｒ Ｒ Ｎ Ａ 基因序列与国外已发现的相应病原基因序列有一定差异，显示其为不同菌株，分别命名为埃立克体 Ｇｚｈ ９８１ 株和 Ｇｚｈ９８２ 株。进一步分析表明，导致我国某养犬基地犬埃立克体病的病原为扁平埃立克体和犬埃立克体。

The causative agent of canine ehrlichiosis had been isolated by animal infection experiment. Then the causative agent was inoculated Beagle pup. Blood was drawn when canine got in fecer, and the genome was extracted. 16S rRNA gene of the agent was amplified, cloned and sequenced. The pathogens were stored in leukocyte including platelet at -80℃, and could infect Beagle pup again. Two different sequences of 16S rRNA gene were amplified and sequenced. There was a discrepancy in the base pair between the 16S rRNA gene of the pathogens and that of Ehrlichia which had been reported respectively. The results showed that the causative agents of canine ehrlichiosis found in China were two species Ehrlichia strain Gzh981 and strain Gzh982. They were regarded as E.platys and E.canis.