Simmons O D, Sobsey M D, Heaney C D, Schaefer F W, Francy D S
School of Public Health, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina 27599-7400, USA.
Appl Environ Microbiol. 2001 Mar;67(3):1123-7. doi: 10.1128/AEM.67.3.1123-1127.2001.
The protozoan parasite Cryptosporidium parvum is known to occur widely in both source and drinking water and has caused waterborne outbreaks of gastroenteritis. To improve monitoring, the U.S. Environmental Protection Agency developed method 1622 for isolation and detection of Cryptosporidium oocysts in water. Method 1622 is performance based and involves filtration, concentration, immunomagnetic separation, fluorescent-antibody staining and 4',6-diamidino-2-phenylindole (DAPI) counterstaining, and microscopic evaluation. The capsule filter system currently recommended for method 1622 was compared to a hollow-fiber ultrafilter system for primary concentration of C. parvum oocysts in seeded reagent water and untreated surface waters. Samples were otherwise processed according to method 1622. Rates of C. parvum oocyst recovery from seeded 10-liter volumes of reagent water in precision and recovery experiments with filter pairs were 42% (standard deviation [SD], 24%) and 46% (SD, 18%) for hollow-fiber ultrafilters and capsule filters, respectively. Mean oocyst recovery rates in experiments testing both filters on seeded surface water samples were 42% (SD, 27%) and 15% (SD, 12%) for hollow-fiber ultrafilters and capsule filters, respectively. Although C. parvum oocysts were recovered from surface waters by using the approved filter of method 1622, the recovery rates were significantly lower and more variable than those from reagent grade water. In contrast, the disposable hollow-fiber ultrafilter system was compatible with subsequent method 1622 processing steps, and it recovered C. parvum oocysts from seeded surface waters with significantly greater efficiency and reliability than the filter suggested for use in the version of method 1622 tested.
已知原生动物寄生虫微小隐孢子虫广泛存在于水源水和饮用水中,并曾引发水源性肠胃炎暴发。为改进监测工作,美国环境保护局制定了方法1622,用于分离和检测水中的隐孢子虫卵囊。方法1622是基于性能的,包括过滤、浓缩、免疫磁分离、荧光抗体染色和4',6-二脒基-2-苯基吲哚(DAPI)复染,以及显微镜评估。将目前方法1622推荐使用的囊式过滤系统与中空纤维超滤系统进行了比较,以对接种的试剂水和未处理地表水进行微小隐孢子虫卵囊的初次浓缩。其他样品按照方法1622进行处理。在使用过滤组件进行的精密度和回收率实验中,对于接种10升试剂水的样品,中空纤维超滤器和囊式过滤器的微小隐孢子虫卵囊回收率分别为42%(标准差[SD],24%)和46%(SD,18%)。在对接种地表水样品同时测试两种过滤器的实验中,中空纤维超滤器和囊式过滤器的平均卵囊回收率分别为42%(SD,27%)和15%(SD,12%)。尽管使用方法1622认可的过滤器能从地表水中回收微小隐孢子虫卵囊,但回收率显著低于试剂级水,且变异性更大。相比之下,一次性中空纤维超滤系统与后续的方法1622处理步骤兼容,并且与测试的方法1622版本中建议使用的过滤器相比,它能以更高的效率和可靠性从接种的地表水中回收微小隐孢子虫卵囊。