云南流产山羊中检出流产嗜性衣原体与Q热立克次氏体混合感染

Detection of Cp .abortus and C. burnetii infection from abortion goat in Yunnan province

建立山羊流产嗜性衣原体与Q热立克次氏体基因组DNA特异扩增检测方法,对云南省河谷地区可疑流产病料时行病原检测并最终证实病原的存在,在此基础上了解病原的进化。获得特异基因产物完成测序验证后递交GenBank,能过BLAST初步分析后调取相关基因,应用DNAstar5.0计算序列间的相似性,计算遗传距离,应用MEGA5.2软件进行比对并构建系统发育树,进行聚类分析。云南流产嗜性衣原体KR778870与有参考序列的衣原体毒株比较其核苷酸同源性均为100%,属同一基因型,与鹦鹉热衣原体同源率最高为94.9%;Q热立克次氏体云南毒株KR697576与纳米比亚毒株CP007555同源性为100%,其次美国毒株为CP00102同源率99.7%,台湾株EU000273为99.4%,同源率最低的毒株EU009657仅为42.6%。通过基因特异性PCR确定流产山羊中检测到流产嗜性衣原体及Q热立克次氏体,流产嗜性衣原体PMP基因无变异,Q热立克次氏体TransⅢ基因则有一定的变异。

The aim of this study was to establish the gene specific PCR method to detect Cp. abor- tus and C. burnteii from abortion goat in Yunnan tropical valley and analyze the target gene varia- tion. The genomic DNA of the samples was extracted and the gene specific PCR products were se- quenced and compared with other reference sequences by DNAstar5. 0 and MEGAS. 2. Results showed that Cp. abortus of Yunnan strain shared 100% nucleotides identify with three reference strains but the homology among Cp. psittaci varied from 42.6% to 100% according to different host. The Yunnan strain shared highly similarity to the strain of Namibia and American. It sug- gested that the PMP gene of Cp. abortus was stably conservative and non-host specific was foun- ded while Tarns Ⅲ gene of C. burnteii was mutated easily in different host.