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目的调查浙江省中部及南部地区巴贝虫动物宿主和传播媒介感染情况,对浙江省确诊的1例人感染巴贝虫的18S rRNA基因序列与Gen Bank中巴贝虫序列进行比对,了解巴贝虫亲缘关系和进化特征。方法采集浙江省确诊的病例居住地以及浙江省中部及南部地区的动物宿主抗凝血样和蜱虫,提取DNA,以巴贝虫18S rRNA基因序列引物进行PCR扩增、测序,并对测序序列进行分析比对,确认感染虫种;在GenBank数据库中查询巴贝虫属、种的18S rRNA基因序列信息,以及不同动物宿主、不同地理来源的田鼠巴贝虫种内18S rRNA序列信息,进行BLAST比对分析,并构建系统进化树。结果采集的261份生物样品中有37份扩增出阳性条带,其中动物宿主抗凝血阳性率为14.2%(32/225),蜱阳性率为13.9%(5/36)。对部分阳性样品进行测序,获得有效测序序列20份,经序列比对分析,共发现田鼠巴贝虫(Babesia microti)5份,均来自鼠类血样;此外,在鼠、牛等宿主/媒介体内还分别检测到肉孢子虫属(Sarcocystis sp.)4份、肝簇虫属(Hepatozoon sp.)2份,球虫(Coccidia sp.)1份、东方泰勒虫(Theileria orientalis)阳性5份、水牛泰勒虫(Theileria buffeli)2份和驽巴贝虫(Babesia caballi)1份。浙江省确诊的人巴贝虫序列与Gen Bank中不同宿主、地理来源田鼠巴贝虫18S rRNA序列的同源性为98.1%~99.8%,与日本、中国云南的人源以及福建、浙江杭州、天台、仙居的鼠源田鼠巴贝虫同属一个分支,而我国的CNMM-2株和新疆1647株与美国GI株和德国Jena株的遗传距离较近。结论在浙江省中部及南部地区的啮齿类小型哺乳动物中发现多份田鼠巴贝虫的自然感染。
Objective To investigate host and vector infection of Babesia in central and southern Zhejiang Province. To compare the 18S rRNA gene sequence of Babeia serogroup 1 infected with human in Zhejiang Province with that in Gen Bank, The relationship and evolutionary characteristics of Bombyx mori. Methods The anticoagulant samples and ticks were collected from the confirmed cases in Zhejiang Province and the animal hosts in central and southern Zhejiang Province. The DNA was extracted and sequenced by PCR with the 18S rRNA gene of Babesia. 18S rRNA gene sequence information of Babesia spp. And species in the GenBank was searched, and 18S rRNA sequence information of Babesia spp. From different animal hosts and from different geographical origins was searched for BLAST ratio Analyze and build phylogenetic trees. Results The positive samples were amplified in 37 out of 261 biological samples. The positive rate of anticoagulant was 14.2% (32/225) in animals and 13.9% (5/36) in ticks. A total of 20 positive samples were sequenced, and 5 copies of Babesia microti were obtained from the blood samples of mice. In addition, in mice and other host / vector Four copies of Sarcocystis sp., Two copies of Hepatozoon sp., One piece of Coccidia sp., Five copies of Theileria orientalis, 2 parts of Theileria buffeli and 1 part of Babesia caballi. The identities of the Babesia parasite in Zhejiang province and those of GenBank in different host and geographical origins were 98.1% ~ 99.8%. The homology of the 18S rRNA sequences with the Japanese, Yunnan, China, Fujian, Zhejiang Hangzhou, Robin Babesia, a mouse-borne Microtus mandarinus on the rooftop and Xianju, belong to one branch, while CNMM-2 and Xinjiang 1647 in China are close to the US GI and Jena strains in Germany. Conclusion Multiple natural rodent Babesia infections were found in small rodents in central and southern Zhejiang Province.