›› 2016, Vol. 43 ›› Issue (4): 1050-1057.doi: 10.16431/j.cnki.1671-7236.2016.04.030

• 疾病防治 • 上一篇    下一篇

牛泰勒虫的分离鉴定及进化分析

刘娟1, 刘军龙1, 刘爱红1, 赵洪喜1, 杨聪山1, 赵帅阳1, 刘光远1, 殷宏1,2, 关贵全1, 罗建勋1   

  1. 1. 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点试验室, 甘肃省动物寄生虫病重点实验室, 兰州 730046;
    2. 江苏省动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009
  • 收稿日期:2015-12-03 出版日期:2016-04-20 发布日期:2016-04-27
  • 通讯作者: 关贵全, 罗建勋 E-mail:guanguiquan@163.com;ljxbn@163.com
  • 作者简介:刘娟(1989-),女,河南驻马店人,硕士生,研究方向:动物寄生虫及其分子生物学,E-mail:cathyliuonline@sina.cn
  • 基金资助:
    国家重点基础研究发展计划(973计划)(2015CB150300);国家肉牛牦牛产业技术体系(NBCIS CARS-38);农业科技创新工程(ASTIP)

The Isolation,Identification and Phylogenetic Analysis of Theileria

LIU Juan1, LIU Jun-long1, LIU Ai-hong1, ZHAO Hong-xi1, YANG Cong-shan1, ZHAO Shuai-yang1, LIU Guang-yuan1, YIN Hong1,2, GUAN Gui-quan1, LUO Jian-xun1   

  1. 1. Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;
    2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease, Yangzhou 225009, China
  • Received:2015-12-03 Online:2016-04-20 Published:2016-04-27

摘要: 牛泰勒虫病是严重危害养牛业可持续发展的血液原虫病,本试验采用血涂片镜检和特异性PCR检测技术,对中国某种牛场的18份血样进行了泰勒虫检测,然后分别用ITS基因通用引物和4种MPSP等位基因特异性引物自阳性样品中扩增出对应的基因,克隆测序后,进行序列比对和进化分析,确定泰勒虫基因型。结果显示,自18份牛血样中检出3份阳性样品,且全为瑟氏泰勒虫感染;3个阳性样品的瑟氏泰勒虫MPSP等位基因扩增结果显示,1个阳性样品为I (Ikeda) 和C (Chitose)型的混合感染,另2个样品为I型单一感染,而均无B (Buffeli)和Thai型;用扩增的MPSP基因测序,构建进化树,确认其感染的瑟氏泰勒虫存在MPSP 1型和2型。这些结果表明,该种牛场存在瑟氏泰勒虫感染,且同时存在2种MPSP等位基因型;MPSP等位基因的复杂性可能使该病的免疫防控更加困难。本试验结果为深入研究瑟氏泰勒虫感染情况及免疫防控提供了数据支持,并为养防一体做好监控。

关键词: 牛泰勒虫病; 血涂片调查; PCR; ITS基因; MPSP等位基因

Abstract: Bovine theileriosis was a kind of hemo-protozoan diseases that seriously hindered the sustainable development of cattle industry.In the present study, the infection of Theileria was detected using microscopic examination and species-specifc PCR of T.annulata, T.sergenti and T.sinensis from 18 blood samples collected from a breeding cattle farm in China.Then ITS and MPSP genes were amplified and cloned using universal primers of ITS and allele-specific primers of 4 major piroplasm surface protein (MPSP) from the positive samples.The sequences were used to make alignment, polymorphism and phylogeny analysis.The results showed that 3 samples were positive for Theileria and the 3 positive samples were all the T.sergenti infection.The amplification of allelic MPSP gene of T.sergenti from the 3 positive samples showed that two of them were single infection by Ikeda-type and another one was co-infection with both Chitose-type and Ikeda-type, while Buffeli-type and Thai-type were not detected.Moreover, on the basis of phylogenetic tree constructed with MPSP gene sequences, types 1 and 2 of MPSP were confirmed to be present in the cattle farm.The results revealed that there were T.sergenti infection in the breeding cattle farm, and these parasites at least with 2 allelic MPSP gene types were present, which indicated that the immune prevention and control of the disease became more complicated.Our research laid foundation of the further study on T.sergenti infection and disease prevention and control.

Key words: bovine theileriosis; microscopic examination; PCR; ITS gene; allelic MPSP gene

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