利用环媒恒温基因扩增法检测生猪刚地弓形虫

On detection of Toxoplasma gondii in swine with loop-mediated isothermal amplification

基于已公布的刚地弓形虫Sag2基因序列设计了1套特异引物,经过条件优化,成功建立了针对刚地弓形虫(简称弓形虫)的环媒恒温基因扩增检测法.根据扩增弓形虫和其他几种病原生物的效果对该方法进行评估.结果表明,该方法只检出致病性弓形虫,特异性良好;检测最低速殖子浓度为4个/μL.对长沙、株洲、湘潭三地生猪血样检测的结果是仔猪、成猪和种母猪弓形虫感染率分别为6.45%、7.69%和15.38%.

Based on the published Sag2 gene sequence of Toxoplasma gondii, a set of oligonucleotide primers were designed. The loop-mediated isothermal amplification （Lamp） method for detection of T.gondii was developed after optimizing the condition. The method established was estimated according to amplification effect of T.gondii and several other pathogenic species, the results showed that the method was of high specificity and its detecting limit was 4 tachyzoites of T.gondii. the blood sample of the swine from cities of Changsha, Zhuzhou and Xiangtan were examined, the finding suggested that the infection rates of Toxoplasma in the piglets, porkers and sows were different, being 6.45%, 7.69% and 15.38% respectively.