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在野生家鼠弓形虫调查中,与传统 DNA 分离相比,磁捕获的效果。

Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice.

机构信息

Department of Pathology and Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic.

Department of Pathology and Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic; Central European Institute of Technology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic.

出版信息

Eur J Protistol. 2014 Feb;50(1):11-5. doi: 10.1016/j.ejop.2013.08.002. Epub 2013 Oct 23.

Abstract

Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 10(4) tachyzoites. Real-time PCR revealed that even 1-5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. The prevalence of T. gondii detected by magnetic capture and qPCR and by commercial isolation and PCR was 1.2% and 0%, respectively. The magnetic capture and quantitative PCR seems to be a highly sensitive and specific diagnostic method for both laboratory research and wild population surveys.

摘要

刚地弓形虫是一种分布广泛的动物源性寄生虫。家鼠(Mus musculus)在寄生虫的生命周期中作为储存宿主起着重要作用。然而,由于宿主体内可能只存在少数组织包囊,因此在家鼠脑中检测到它们的存在受到限制。为了提高诊断效果,我们测试了一种新的用于检测鼠脑中弓形虫的方案,并将该技术与使用商业 DNA 分离试剂盒的基于聚合酶链反应(PCR)的标准方案进行了比较。我们测试了用磁场从整个宿主大脑中分离弓形虫 DNA 的效果,实验向实验室鼠的大脑样本中添加了 1 到 10(4)个速殖子。实时 PCR 显示,即使在磁场捕获后,也能检测到 1-5 个速殖子。此外,该方法还适用于定量检测大脑中超过 10 个速殖子等价物的寄生虫数量。为了在野生鼠中评估这两种技术,我们使用了一个由 243 个个体组成的数据集。通过磁场捕获和 qPCR 以及商业分离和 PCR 检测到的弓形虫的流行率分别为 1.2%和 0%。因此,磁场捕获和定量 PCR 似乎是一种对实验室研究和野生种群调查都具有高度敏感性和特异性的诊断方法。

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