湖南省牛无浆体虫株分子生物学鉴定

Study on Molecular Biology Identification of Bovine Anaplasma Isolates in Hunan

用原虫16S rRNA基因序列通用引物对湖南5个地区牛无浆体感染的阳性血液样品进行PCR扩增,经测序和序列拼接,获得5个无浆体样品16S rRNA基因的部分序列,应用分子生物学软件进行分析,并根据所获得序列的保守区间设计特异性引物,进行PCR诊断方法的探讨性研究。结果表明,在通用引物下,5个地区无浆体感染的阳性血液样品均获得1425bp大小的虫体16S rRNA基因条带;测序后5个无浆体16S rRNA序列同源性均在97%～98%。证明5个分离虫株初步鉴定为牛边缘无浆体。

The 16 S RNA gene sequence of Anaplasma was amplificated from positive blood in cattle with Anaplasrna infection in five different districts of Hunan with the universal primers of protozoa. PCR products were partially sequenced and spliced, and ]6sRNA gene partial sequence of Anaplasma isolated from different districts in Hunan were analyzed or identified via molecular biological software. At last, a specific PCR diagnosis method on Anaplasma was researched with a pair of specific designed primers in the conservative interval of such acquired se- quence in the study. Results indicated that a 1 425 bp gene strap of Anaplasma ]6 S RAN was obtained in all of the positive blood samples in cattle via PCR. Sequence homology of 16 S rRNA among 5 isolates of Anaplasma from different districts of Hunan and other 10 isolates of Anaplasma marginale on GenBank were 97%-98%.