原生动物寄生虫微小隐孢子虫的水传播仍然是一个问题 严重影响免疫功能低下人群的重要疾病来源。结束 过去十年，中国在解决隐孢子虫问题方面取得了很大进展 水临时强化地表水处理规则（IESWTR）和长期 1美国环境保护局（USEPA）颁布了强化地表水处理规则（LT1ESWTR），以 降低隐孢子虫病的风险。有待解决的长期问题2增强了地表水 处理规则（LT2ESWTR）将要求对来源不良的公用事业进行额外处理 水质。此外，许多公用事业公司，包括美国的所有公用事业子公司 水，已经加入了一个自愿的安全饮用水伙伴关系，以进一步加强运营 改进隐孢子虫卵囊治疗的标准。 免疫荧光抗体（IFA）试验已用于检测小鼠隐孢子虫卵囊 经过过滤的饮用水，但该检测方法有很多局限性。IFA分析 无法确定饮用水中检测到的卵囊的公共卫生意义。作为一个 结果，美国研究人员开发了一种检测活的、感染性的病毒的专利测试 隐孢子虫卵囊在水中。 这种方法被称为“细胞培养，聚合酶链” “反应”（CC-PCR）检测是准确的，因为它测量 隐孢子虫能够在人类肠道细胞中生长，分离出感染性病原体 从水样中提取卵囊。 对过滤后的饮用水样本应用CC-PCR检测有助于确定 水务公司面临的核心问题:当前的水处理技术是否充分 控制水中隐孢子虫的风险？目前的研究就是为了回答这个问题 提出问题并提供控制水中隐孢子虫的策略。每月从82个常规地表水收集工厂废水样品 处理厂位于14个州，共分析了1690 100-L成品水样 通过CC-PCR技术。成品水样用2%的溶液中和 硫代硫酸钠（密苏里州圣路易斯西格玛市）使用直列DEMA注射器， MO）以10.0 ml/min的流速采集隐孢子虫样本 方法1622（美国环保局，1998年），使用Envirochek（TM）采样胶囊（Pall Life Sciences， 并以2.0升/分钟的流速过滤100升成品水。 用细胞培养PCR（CC）检测感染性隐孢子虫卵囊- PCR）检测 （Di Giovanni等人，1999年）。对于阳性对照组，来自 Harley Moon品系（NADC-USDA、Ames、IA），牛基因型；年龄不超过四周的患者被排除在外 购买自斯特林寄生虫学实验室，亚利桑那大学，I（Tucson，AZ）。到 在确定隐孢子虫分离株的基因型后，利用PCR技术克隆了hsp70扩增子 TOPO-TA克隆试剂盒（Invitrogen；加利福尼亚州卡尔斯巴德）符合制造商的要求 说明书排除实验室阳性对照样本的可能性（Harley Moon 菌株卵囊）可能在加工过程中污染了成品饮用水样本， 最初的hsp70 PCR反应，在-80℃或-20℃下冷冻2-24小时 如Strong等人（2000年）所述，对gp60标记物进行了几个月的复查。包括27个参考文献、表格和图表。

Waterborne transmission of the protozoan parasite Cryptosporidium parvum remains a significant source of disease with severe consequences for immunocompromised people. Over the past decade much progress has been made in addressing the issue of Cryptosporidium in water. Both the Interim Enhanced Surface Water Treatment Rule (IESWTR) and the Long Term 1 Enhanced Surface Water Treatment Rule (LT1ESWTR) were promulgated by the US Environmental Protection Agency (USEPA) to reduce the risk of cryptosporidiosis. The pending Long Term 2 Enhanced Surface Water Treatment Rule (LT2ESWTR) will require additional treatment for utilities with poor source water quality. In addition, many utilities, including all of the utility subsidiaries of American Water, have joined a voluntary Partnership for Safe Drinking Water to further tighten operating standards to improve the treatment of Cryptosporidium oocysts. The immunofluorescent antibody (IFA) test has been used to detect Cryptosporidium oocysts in filtered drinking water but the assay has a number of limitations. The IFA assay cannot determine the public health significance of oocysts detected in drinking water. As a result, American researchers developed a patented test for detecting live, infectious Cryptosporidium oocysts in water. The method called the "cell culture, polymerase chain reaction" (CC-PCR) test is accurate and precise because it measures the DNA of Cryptosporidium that are able to grow in human intestinal cells after the isolation of infectious oocysts from water samples. Application of the CC-PCR assay to filtered drinking water samples can help determine the central question facing water utilities: do current water treatment techniques adequately control the risks of Cryptosporidium in water? The current study was initiated to answer this question and provide a strategy for control of Cryptosporidium in water. Monthly finished plant effluent samples were collected from 82 conventional surface water treatment plants, located in 14 states, for a total of 1690 100-L finished water samples analyzed by the CC-PCR technique. Finished water samples were neutralized with a solution of 2% sodium thiosulfate (Sigma, St. Louis, MO) using an inline DEMA injector (DEMA, St. Louis, MO) at a flow rate of 10.0 ml/min. Cryptosporidium samples were collected according to Method 1622 (USEPA, 1998) using the Envirochek(TM) sampling capsule (Pall Life Sciences, Ann Harbor, MI) and filtration of 100 L of finished water at a flow rate of 2.0 L/minute. Infectious Cryptosporidium oocysts were detected using the cell culture PCR (CC-PCR) assay (Di Giovanni et al. 1999). For positive controls, live Cryptosporidium parvum oocysts from the Harley-Moon strain (NADC-USDA, Ames, IA), bovine genotype; no older than four weeks were purchased from the Sterling Parasitology Laboratory, University of Arizona, I(Tucson, AZ). To determine the genotype of the Cryptosporidium isolates, the hsp70 amplicon was cloned using the TOPO TA Cloning Kit (Invitrogen; Carlsbad, CA) in accordance with the manufacturer's instructions. To rule out the possibility that laboratory positive control samples (Harley-Moon strain oocysts) could have contaminated finished drinking water samples during processing, original hsp70 PCR reactions, which had been kept frozen (at -80C or -20C) for two to twenty-four months were reexamined for the gp60 marker as described by Strong et al. (2000). Includes 27 references, tables, figures.