Environmental Microbial and Food Safety Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Building 173, BARC-East, 10300 Baltimore Avenue, Beltsville, MD 20705, USA.
Vet Parasitol. 2009 Dec 3;166(1-2):32-7. doi: 10.1016/j.vetpar.2009.07.039. Epub 2009 Aug 3.
Four species of Cryptosporidium are routinely found in cattle: Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae, and Cryptosporidium andersoni. It is important to determine the species of Cryptosporidium in infected cattle because C. parvum is the only serious pathogen for humans as well as cattle. Identification of Cryptosporidium species and genotypes currently relies on molecular methods such as polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) or gene sequencing. Incorporation of these techniques in a routine veterinary diagnostic laboratory is cost prohibitive. As such, their applications are limited primarily to research and a few public health laboratories. To overcome this problem, a multiplex PCR assay was developed for simultaneously detecting the 4 species of Cryptosporidium that commonly infect cattle. This assay specifically identifies Cryptosporidium oocysts present in cattle feces, improves the detection of mixed infections, reduces the time and cost relative to current sequencing methods, and further demonstrates the shortcomings of sequencing as the definitive method for identification when analyzing samples containing mixed infections.
微小隐孢子虫、牛隐孢子虫、瑞安隐孢子虫和犬隐孢子虫。确定感染牛的隐孢子虫种类非常重要，因为微小隐孢子虫是唯一对人类和牛都严重的病原体。隐孢子虫种和基因型的鉴定目前依赖于分子方法，如聚合酶链反应（PCR），然后是限制性片段长度多态性（RFLP）或基因测序。将这些技术纳入常规兽医诊断实验室的成本过高。因此，它们的应用主要限于研究和少数公共卫生实验室。为了解决这个问题，开发了一种多重 PCR 检测方法，用于同时检测通常感染牛的 4 种隐孢子虫。该检测方法专门鉴定存在于牛粪便中的隐孢子虫卵囊，提高了混合感染的检测率，与当前的测序方法相比，缩短了时间和成本，进一步证明了在分析包含混合感染的样本时，测序作为鉴定的确定方法存在缺陷。
