评估用于检测水中贾第虫包囊和隐孢子虫卵囊的免疫荧光程序。
Evaluation of the immunofluorescence procedure for detection of Giardia cysts and Cryptosporidium oocysts in water.
作者信息
LeChevallier M W, Norton W D, Siegel J E, Abbaszadegan M
机构信息
American Water Works Service Company, Inc., Voorhees, New Jersey 08043, USA.
出版信息
Appl Environ Microbiol. 1995 Feb;61(2):690-7. doi: 10.1128/aem.61.2.690-697.1995.
The accurate determination of the presence of Giardia cysts and Cryptosporidium oocysts in surface waters requires a reliable method for the detection and enumeration of these pathogenic organisms. Published methods have usually reported recovery efficiencies of less than 50% for both cysts and oocysts. Typically, the losses are greater for Cryptosporidium oocysts than they are for Giardia cysts. The purpose of this study was to examine procedures used for sample collection, elution, concentration, and clarification to determine when losses of cysts and oocysts occurred during processing. The results showed that major losses of cysts and oocysts occurred during centrifugation and clarification. Depending on the centrifugation force, oocyst losses of as high as 30% occurred for each centrifugation step. A 1.15-specific-gravity Percoll-sucrose gradient was needed to optimize recovery of oocysts from natural water samples. Minor improvements in the procedure could be accomplished by selecting a filter other than the recommended 1-micron-pore-size (nominal-porosity) polypropylene filter.
准确测定地表水中贾第虫包囊和隐孢子虫卵囊的存在，需要一种可靠的方法来检测和计数这些致病生物体。已发表的方法通常报告包囊和卵囊的回收率均低于50%。通常，隐孢子虫卵囊的损失比贾第虫包囊的损失更大。本研究的目的是检查用于样品采集、洗脱、浓缩和澄清的程序，以确定在处理过程中何时发生包囊和卵囊的损失。结果表明，包囊和卵囊的主要损失发生在离心和澄清过程中。根据离心力的不同，每个离心步骤的卵囊损失高达30%。需要1.15比重的Percoll-蔗糖梯度来优化从天然水样中回收卵囊。通过选择推荐的1微米孔径（标称孔隙率）聚丙烯滤膜以外的滤膜，可以对该程序进行微小改进。
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