本文介绍了一种快速简便的检测微小隐孢子虫的聚合酶链反应（PCR）方法。该方案使用片剂形式制备的PCR试剂和针对微小隐孢子虫的PCR引物。PCR试剂片的使用减少了制备PCR试剂鸡尾酒所需的时间和人力，提高了重现性，并降低了污染的可能性。每个PCR检测还包含一个耐热DNA插入荧光染料。PCR产物的终点检测使用荧光计进行，荧光计可容纳PCR管，提供不需要对PCR产物进行凝胶电泳分析的封闭管分析。由于包含已完成反应的试管无需打开，因此PCR产物污染实验室的可能性大大降低。 目前，使用荧光计进行终点检测时，使用纯化的卵囊储备和冻融循环制备的模板DNA制备的检测限为五个卵囊。对于含有免疫磁分离（IMS）纯化环境原水浓缩物（相当于每PCR 0.5 mL填充颗粒）的种子PCR检测，获得了10个卵囊的检测限。该试剂盒的未来开发可能会结合基于荧光探针的技术来检测特定的PCR产物，从而可以检测单个微小隐球菌卵囊。

This paper describes a rapid and simple polymerase chain reaction (PCR) protocol for the detection of Cryptosporidium parvum. This protocol uses PCR reagents which have been prepared in tablet form and PCR primers specific for Cryptosporidium parvum. The use of PCR reagent tablets reduces time and labor required to prepare PCR reagent cocktails, increases reproducibility, and decreases the potential for contamination. Each PCR assay also contains a thermostable DNA intercalating fluorescent dye. Endpoint detection of PCR product is performed using a fluorometer which accommodates PCR tubes, providing a closed tube assay which does not require gel electrophoresis analysis of PCR products. Since the tubes containing the completed reactions do not need to be opened the chance of laboratory contamination with PCR product is greatly reduced. Endpoint detection using the fluorometer currently has a detection limit of five oocysts using a purified oocyst stock and template DNA preparation by freeze-thaw cycling. A detection limit of ten oocysts was obtained for seeded PCR assays containing immunomagnetic separation (IMS) purified environmental raw water concentrates (equivalent to 0.5 mL packed pellet per PCR). Future development of the kit may incorporate fluorescent probe-based technology for the detection of specific PCR product which may allow the detection of a single C. parvum oocyst.