分子鉴定2种猪食道口线虫熔解曲线分析试验的建立  

Establishment of Melting Curve Analysis Assay for Molecular Identification of Oesophagostomum spp. from Pigs

作　　者：舒丽[1,2] 程田[1] 翁亚彪[1] 林瑞庆[1] 邹尚书 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]四川省富顺县永年镇兽医站,四川富顺643200

出　　处：《中国畜牧兽医》2013年第9期70-74,共5页China Animal Husbandry & Veterinary Medicine

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(IRT0723);广东省高等学校大学生创新实验项目

摘　　要：基于猪有齿食道口线虫和四棘食道口线虫IGS rDNA序列的遗传变异情况,于种内保守、种间差异明显的序列片段设计1对引物,建立了区分2种猪食道口线虫的熔解曲线分子鉴定方法。研究结果显示,该方法可通过熔解曲线差异而准确鉴定猪食道口线虫2个种,与其他线虫和吸虫无交叉反应,特异性良好;能检测猪食道口线虫单虫卵样品和FTA卡片的DNA模板。该研究为猪食道口线虫种类准确鉴定提供了一种新的准确、敏感、特异且快速的分子手段。Based on the IGS rDNA sequences of O.dentatum and O.quadrispinulatum, a pair of primers （MCF/MCR） was designed for developing a melting curve analysis assay for the identification and differentiation of two Oesophagostomum species. This assay allowed the specific identification and differentiation of O.dentatum and O.quadrispinulatum by melting curve analysis, with no amplicons being amplified from heterogenerous DNA samples. DNA sequencing confirmed the identity of the amplified products. The single egg of Oesophagostomum spp. and DNA extracted using What-man FTA card could also be detected by this assay. These findings showed that the melting curve analysis assay established was accurate, sensitive, specific and rapid, allowing the detection and identification of O.dentatum and O.quadrispinulatum from pigs.

关 键 词：有齿食道口线虫 四棘食道口线虫 熔解曲线分析 分子鉴定 

分 类 号：S852.72[农业科学—基础兽医学]