河南农业科学 ›› 2022, Vol. 51 ›› Issue (12): 147-152.DOI: 10.15933/j.cnki.1004-3268.2022.12.017

• 畜牧·兽医 • 上一篇    下一篇

绵羊源十二指肠贾第虫和隐孢子虫的分子鉴定

睢攀博1,2,徐菲菲1,梁冠达1,杜海利1,郎家抒1,李俊强1   

  1. (1.河南农业大学动物医学院/农业农村部禽类产品质量安全控制重点实验室,河南 郑州 450046;2.河南省滑县动物卫生监督所,河南 滑县 456400)
  • 收稿日期:2022-05-06 出版日期:2022-12-15 发布日期:2023-01-17
  • 通讯作者: 李俊强(1987-),男,河南林州人,副教授,主要从事人兽共患寄生虫生物学研究。E-mail:lijunqiangcool@126.com
  • 作者简介:睢攀博(1986-),男,河南滑县人,兽医师,硕士,主要从事兽医行政执法工作。E-mail:1768576337@qq.com
  • 基金资助:
    国家自然科学基金项目(32102689);河南农业大学青年英才基金项目(30501055)

Molecular Identification of Giardia duodenalis and Cryptosporidium spp.from Sheep

SUI Panbo1,2,XU Feifei1,LIANG Guanda1,DU Haili1,LANG Jiashu1,LI Junqiang1   

  1. (1.College of Veterinary Medicine,Henan Agricultural University/Key Laboratory of Quality and Safety Control of Poultry Products(Zhengzhou),Ministry of Agriculture and Rural Affairs,Zhengzhou 450046,China;2.Huaxian County Animal Health Supervision Institute of Henan Province,Huaxian 456400,China)
  • Received:2022-05-06 Published:2022-12-15 Online:2023-01-17

摘要: 为了解河南省安阳市绵羊源贾第虫和隐孢子虫的分子特性,对基于卢戈氏碘液染色法和饱和蔗糖溶液漂浮法检查获得的16份绵羊源贾第虫和3份隐孢子虫分离株分别进行DNA提取,并基于16SrRNA基因位点和18S rRNA基因位点进行巢式PCR扩增,获得阳性扩增产物,再进行测序鉴定和系统发育进化分析。结果显示,PCR成功扩增绵羊源贾第虫和隐孢子虫的特异性基因位点。基于对上述2种肠道原虫特异性基因位点的分子序列分析,将贾第虫鉴定为十二指肠贾第虫集聚体E(93.75%,15/16)和集聚体A(6.25%,1/16);将隐孢子虫鉴定为泛在隐孢子虫(66.67%,2/3)和猪隐孢子虫(33.33%,1/3)。

关键词: 绵羊, 十二指肠贾第虫, 隐孢子虫, 分子鉴定, 安阳市

Abstract: To investigate the molecular characteristics of Giardia and Cryptosporidium from sheep in Anyang area,Henan Province,microscopy positive fecal samples based on Lugol’s iodine staining techniques and Sheather’s sugar flotation were obtained,followed by DNA extraction from sixteen Giardia positive samples and three Cryptosporidium positive samples.The nested PCR amplification based on 16S rRNA locus and 18S rRNA locus was performed,respectively.The positive amplicons were sequenced,followed by phylogenetic and evolutionary analysis.The results indicated that the specific gene loci of Giardia and Cryptosporidium from sheep were successfully amplified by PCR.Molecular characteristics analysis of the specific loci of the two intestinal protozoans showed that the Giardia strains were identified as G.duodenalis assemblage E(93.75%,15/16)and assemblage A(6.25%,1/16).And the Cryptosporidium strains were identified as C.ubiquium(66.67%,2/3)and C.suis(33.33%,1/3).

Key words: Sheep, Giardia duodenalis, Cryptosporidium spp., Molecular identification, Anyang

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