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首页> 外文期刊>Journal of Clinical Microbiology >Development of a Real-Time PCR Assay for Detection of Toxoplasma gondii in Pig and Mouse Tissues
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Development of a Real-Time PCR Assay for Detection of Toxoplasma gondii in Pig and Mouse Tissues

机译：猪和小鼠组织中弓形虫实时PCR检测方法的开发

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摘要

A highly sensitive and specific method has been developed to reproducibly detect and quantitate Toxoplasma gondii burden in animal tissue samples using T. gondii ITS1-derived primers and a fluorogenic probe via real-time PCR. Assay specificity was confirmed against a panel of DNA samples from T. gondiiand other common protozoa as well as host animal tissue. This Toxo TaqMan assay was able to detect as little as 0.1 pg of T. gondii genomic DNA, which is equivalent to 1 T. gondii bradyzoite, and has a dynamic range of detection of from 100 ng to 100 fg of T. gondii DNA. Tissues from experimentally infected mice and pigs as well as bradyzoite-spiked pig muscle samples were used to test and standardize this technique. Positive signals were obtained with T. gondii parasite concentrations ranging from 4 to 3.7 × 105 parasites per g of spiked pig tissue, with excellent linearity (R 2 = 0.9776). All T. gondii-infected animals were correctly identified by this technique. Results indicate that this assay is applicable to swine carcasses and commercial pig products, is compatible with automation technology for potential slaughterhouse use, and will enable scientists to diagnose and quantitate T. gondii in animal tissues.
机译：已开发出一种高度灵敏，特异的方法，使用 T可重复检测和定量动物组织样品中的弓形虫弓形虫负担。实时聚合酶链反应（ITS）衍生的弓形虫引物和荧光探针。针对来自 T的一组DNA样品确认了测定特异性。刚地犬和其他常见的原生动物以及宿主动物组织。这种Toxo TaqMan分析方法能够检测到0.1 pg的Tem。 gondii 基因组DNA，相当于1个 T。 gondii 缓殖子，动态检测范围为100 ng至100 fg的 T。刚地 DNA。实验感染的小鼠和猪的组织以及缓殖子掺入的猪肌肉样品均用于测试和标准化该技术。用 T获得正信号。每克加标的猪组织中刚德虫的寄生虫浓度为4至3.7×10 5 寄生虫，线性极佳（ R 2 = 0.9776）。所有 T。用这种技术正确地鉴定了刚地犬感染的动物。结果表明，该测定法适用于猪的尸体和商品猪产品，与自动化技术兼容，可用于潜在的屠宰场，并将使科学家能够诊断和定量 T。动物组织中的刚地。

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