提高诊断的特异性 采用实时定量PCR（qPCR）检测水中隐孢子虫 用于检测和定量环境中的隐孢子虫卵囊 样品；然而，这些方法具有不同程度的特异性和敏感性 并不是人类梭状芽胞杆菌或微小梭状芽胞杆菌特有的。因此，本研究的目标 该项目旨在改进现有的qPCR检测方法，以特异性检测微小隐球菌，C。 人隐孢子虫，或所有隐孢子虫属。 该项目实验方法的第一个目标是开发三种qPCR分析。第一个 是一种检测所有隐孢子虫物种的“泛隐”qPCR检测方法，而第二种 第三种方法分别特异性检测微小隐球菌或人隐球菌卵囊。 初步结果表明，pan-Crypto-qPCR法可以方便地检测基因组 人传染性隐孢子虫和动物性隐孢子虫的DNA提取物。 比如C。 犬类、猫科动物、人科动物、马利亚科动物、鼠科动物和微小类动物。附加的 实验进一步表明，泛加密qPCR可以检测到低至1的水平 卵囊在试剂级水中。对于开发的其他qPCR分析，C.parvum 并且只检测到猫的C。 meleagridis，或C.muris。类似地，人型梭菌特异性qPCR检测仅检测到C。 人是卵囊，而不是犬类梭菌、猫类梭菌、马类梭菌、鼠类梭菌或微小梭菌。 进一步评估所有检测限和特异性的附加实验 本文讨论了三种检测环境样品中添加的卵囊的方法。本项目实验方法的第二个目标是确定qPCR结果的变化 使用不同的实时PCR机器生成。到目前为止，有三个是真实的- 时间PCR 比较了ABI 7000、ABI 7900和罗氏LC480智能自行车。 初步结果表明，所有三台受试机器都能检测到低浓度 模板DNA，相当于一个卵囊，未观察到显著差异。 其他实时PCR机器的性能结果也将在发布时展示 随时可用。包括11篇参考文献，仅为摘要。

To improve the specificity of detecting Cryptosporidium in water, quantitative real-time PCR (qPCR) assays were developed to detect and quantitate Cryptosporidium spp. oocysts in environmental samples; however, these approaches have varying degrees of specificity and sensitivity and were not specific to C. hominis or C. parvum. Therefore, the goal of this project was to improve upon current qPCR assays to specifically detect C. parvum, C. hominis, or all Cryptosporidium spp. The first goal of the experimental approach of this project was the development of three qPCR assays. The first is a "pan-Crypto" qPCR assay that detects all Cryptosporidium species, while the second and third assays specifically detect C. parvum or C. hominis oocysts, respectively. Preliminary results showed that the pan-Crypto qPCR assay can easily detect genomic DNA extracts from human infectious and animal forms of Cryptosporidium spp. like C. canis, C. felis, C. hominis, C. meleagridis, C. muris and C. parvum. Additional experiments further revealed that the pan-Crypto qPCR can detect levels as low as one oocyst in reagent grade water. For the other qPCR assays developed, the C. parvum specific qPCR only detected C. parvum oocysts and not C. canis, C. felis, C. hominis, C. meleagridis, or C. muris. Similarly, the C. hominis specific qPCR assay only detected C. hominis oocysts and not C. canis, C. felis, C. meleagridis, C. muris, or, C. parvum. Additional experiments that further evaluate limits of detection and specificities of all three assays in detecting oocysts spiked in environmental samples are discussed. The second goal of the experimental approach of this project was to determine the variations of qPCR results generated from using different real-time PCR machines. To date, three real-time PCR machines, ABI 7000, ABI 7900, and Roche LC 480 Smart Cycler, were compared. Preliminary results indicated that all three machines tested were able to detect low levels of template DNA, equivalent to one oocyst, and no significant differences were observed. Performance results from other real-time PCR machines will also be presented as they become available. Includes 11 references, abstract only.