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中国中部河南省奶牛新孢子虫的血清流行率及首次多位点微卫星基因分型

Seroprevalence and first multilocus microsatellite genotyping of Neospora caninum in dairy cattle in Henan, central China.

作者信息

Qian Weifeng, Wang Tianqi, Yan Wenchao, Zhang Min, Han Lifang, Xue Rui, Song Shaofu, Lv Chaochao

机构信息

College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China.

College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China.

出版信息

Vet Parasitol. 2017 Sep 15;244:81-84. doi: 10.1016/j.vetpar.2017.07.022. Epub 2017 Jul 25.

Abstract

Neospora caninum is one of the important causes of abortion in cattle worldwide, and losses due to neosporosis to the cattle industry are considerable. However, the knowledge of genetic characterization of this parasite is limited. The aim of the present study is to determine the prevalence and genetic characterization of N. caninum from dairy cows in Henan Province, central China. A total of 510 blood samples and 7 aborted fetuses were collected from 8 dairy farms in Henan Province. Serum antibodies to N. caninum were examined by ELISA using a recombinant tNcSRS2 protein as the coating antigen. The overall seroprevalence of N. caninum in dairy cows was 41.2% (210/510). The seropositivity rate of N. caninum in aborting cows (49.3%) was statistically significant higher than that (29.3%) in non-aborting cows (p<0.05) with an odds ratio of 2.44 (95% CI, 1.61-3.41). Statistical association was also found between farm type and the seropositivity rate of N. caninum infection in cows (p<0.01).N. caninum DNA was detected from 6 of 396 blood samples (1.5%) and 4 of 7 aborted fetuses by nested PCR based on NC5 gene, and the 10N. caninum positive DNA samples were further analyzed by multilocus microsatellite (MS) genotyping for MS4, MS5, MS6A, MS7, MS8, MS10, and MS12. Only 2 samples were successfully genotyped at all genetic loci, and two unique profiles including two novel allelic patterns were identified. To our knowledge, this study is the first report of genetic characterization of N. caninum isolates from naturally infected dairy cows based on multilocus microsatellites (more than 2 loci) in China.

摘要

犬新孢子虫是全球范围内牛流产的重要病因之一,新孢子虫病给养牛业造成的损失相当大。然而,关于这种寄生虫遗传特征的了解有限。本研究的目的是确定中国中部河南省奶牛中犬新孢子虫的流行情况和遗传特征。从河南省8个奶牛场共采集了510份血液样本和7个流产胎儿。以重组tNcSRS2蛋白为包被抗原,采用ELISA法检测犬新孢子虫血清抗体。奶牛中犬新孢子虫的总体血清阳性率为41.2%(210/510)。流产奶牛中犬新孢子虫的血清阳性率(49.3%)显著高于未流产奶牛(29.3%)(p<0.05),优势比为2.44(95%CI,1.61 - 3.41)。还发现农场类型与奶牛中犬新孢子虫感染的血清阳性率之间存在统计学关联(p<0.01)。基于NC5基因的巢式PCR从396份血液样本中的6份(1.5%)和7个流产胎儿中的4份检测到犬新孢子虫DNA,对10份犬新孢子虫阳性DNA样本进一步进行多位点微卫星(MS)基因分型,检测MS4、MS5、MS6A、MS7、MS8、MS10和MS12。仅2份样本在所有基因位点成功进行了基因分型,并鉴定出两种独特的图谱,包括两种新的等位基因模式。据我们所知,本研究是中国首次基于多位点微卫星(超过2个位点)对自然感染奶牛的犬新孢子虫分离株进行遗传特征分析的报告。

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