SYBR Green Ⅰ荧光定量PCR鉴定简单异尖线虫方法的建立  

Establishment of Anisakis Simplex by SYBR Green Ⅰ Real-Time Fluorescent Quantitative PCR

作　　者：龚艳清[1] 郑洋妹[2] 陈信忠[1] 

机构地区：[1]厦门出入境检验检疫局,福建厦门361026 [2]福建农林大学动物科学学院

出　　处：《检验检疫学刊》2010年第6期28-31,共4页Journal of Inspection and Quarantine

摘　　要：根据GenBank发表的简单异尖线虫保守的ITS-2基因序列设计一对引物,用以扩增简单异尖线虫114 bp基因片段。用简单异尖线虫的重组质粒(AS-ITS-pGM)为模板建立SYBR Green I荧光定量PCR检测简单异尖线虫的方法,并用该方法对经PCR-RFLP鉴定为简单异尖线虫的样品进行鉴定。结果表明,本研究建立的简单异尖线虫SYBR Green I荧光定量PCR方法特异性强、敏感性高,稳定性好,可用于简单异尖线虫的鉴定。According the internal transcribed spacer（ITS） gene sequences of the nematode of Anisakis simplex,a set of specific primers were designed to amplify the special DNA sequence by the technique of fluorescent quantitative PCR based on SYBR Green Ⅰ.A 114 bp fragment had been amplified from A.simplex and used to construct the recombinant plasmid of AS-ITS-pGM.The plasmid was used as a template that amplified in the fluorescent quantitative PCR to detect A.simplex.The verify experiment had proved that the established method had high sensitivity and specificity,good stability,and can be used to identify the nematode of A.simplex.

关 键 词：SYBRGreenI 荧光定量PCR 简单异尖线虫 

分 类 号：S852.731[农业科学—基础兽医学]