对环境水样中自然产生的微小隐球菌卵囊进行免疫磁性分离（IMS），然后使用集成细胞培养PCR（CC-PCR）进行感染性测定的策略进行了现场试验。对来自美国25个地点的100多个原水和过滤反冲洗水抓取样本进行了分析。卵囊种子原水和过滤反冲洗水样也用于评估CC-PCR方案在不同水质基质下的回收效率和性能。抓取的样品通过离心浓缩，浓缩物通过IMS分离和纯化。 含有1%胰蛋白酶的酸化汉克平衡盐溶液（HBSS）用于从IMS珠中分离捕获的卵囊，因为制造商建议的0.1 N HC1分离可能会对卵囊的活力产生不利影响。在96孔微量滴定板中进行IMS纯化卵囊的体外HCT-8细胞培养，并使用针对微小隐球菌的PCR引物检测感染细胞。CC-PCR在6个原水和9个过滤反冲洗水样中检测到传染性微小隐孢子虫。所有CC-PCR阳性样本均通过PCR产物的克隆和DNA序列分析得到确认。

Field testing of a strategy using immunomagnetic separation (IMS) of naturally occurring C. parvum oocysts from environmental water samples followed by infectivity determination using integrated cell culture-PCR (CC-PCR) was initiated. Over 100 each raw source water and filter backwash water grab samples from twenty-five sites in the U.S. have been analyzed. Oocyst seeded raw and filter backwash water samples were also used to evaluate recovery efficiencies and performance of the CC-PCR protocol with different water quality matrices. The grab samples were concentrated by centrifugation, concentrates split and purified by IMS. An acidified Hank's balanced salt solution (HBSS) containing 1% trypsin was used for the dissociation of captured oocysts from the IMS beads since the manufacturer recommended 0.1 N HC1 dissociation may adversely affect oocyst viability. In vitro HCT-8 cell culture of IMS purified oocysts was performed in 96-well microtiter plates and infected cells were detected using PCR primers specific for C. parvum. CC-PCR has detected infectious Cryptosporidium parvum in 6 raw and 9 filter backwash water samples. All CC-PCR positive samples were confirmed by cloning and DNA sequence analysis of the PCR products.