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基于弓形虫529-bp重复序列巢式PCR诊断方法的建立

Establishment of nested PCR based on 529-bp repetitive sequence for diagnosis of Toxoplasma gondii

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【作者】 张莹光曹利利徐丹宋鑫帅魏峰许越王巍杨桂连刘全

【Author】 ZHANG Ying-guang1,2,CAO Li-li2,3,XU Dan1,2,SONG Xin-shuai1,2,WEI Feng1,XU Yue1,WANG Wei2,YANG Gui-lian4,LIU Quan2 (1.College of Life Science,Jilin Agricultural University,Changchun 130118,China;2.Key Laboratory of Jilin Province for Zoonosis Prevention and Control,Institute of Military Veterinary,Academy of Military Medical Sciences,Changchun 130122,China;3.Jilin Institute of Animal Science and Veterinary Medicine,Changchun 130062,China;4.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)

【机构】 吉林农业大学生命科学学院军事医学科学院军事兽医研究所吉林省人畜共患病预防与控制重点实验室吉林省畜牧兽医科学研究院吉林农业大学动物科学技术学院

【摘要】 根据GenBank已发表的弓形虫529-bp重复序列(AF146527)设计巢式PCR引物,建立巢式PCR检测方法。结果表明,该方法能扩增出427bp的片段,敏感性试验表明该方法可以检测出0.1pg的弓形虫基因组DNA,敏感性是常规PCR的100倍。巢式PCR方法对蜥蜴利什曼原虫、隐孢子虫、细粒棘球绦虫基因组扩增无条带,特异性强。样品检测符合率达100%。巢式PCR方法的建立为弓形虫病的诊断及流行病学调查提供技术支持。

【Abstract】 The nested PCR primers were designed according to GenBank published 529-bp repetitive DNA sequence(AF146527) in Toxoplasma gondii,we have established the nested PCR.The product of the nested PCR was about 427 bp and its sensitivity was 100 times than that of conventional PCR,which could detect the genomic DNA as low as 0.1 pg.Detection of Leishmania tarentolae,Cryptosporidium parvum and Echinococcus granulosus genome DNA by nested PCR had no fragment.The diagnosis coincidence rate is 100%.The nested PCR method is dependable for diagnosis of Toxoplasma gondii.

【基金】 国家自然科学基金资助项目(30972178,31072127,31001057);国家“863”计划资助项目(2011AA10A215)
  • 【文献出处】 中国兽医学报 ,Chinese Journal of Veterinary Science , 编辑部邮箱 ,2013年02期
  • 【分类号】S855.9
  • 【被引频次】9
  • 【下载频次】275
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