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目的建立水源水中贾第鞭毛虫和隐孢子虫的滤膜溶解-免疫磁分离-荧光染色检测方法。方法水样经混合乙酸纤维膜过滤后,溶解于丙酮,离心浓缩,用免疫磁珠分选浓缩产物中的贾第鞭毛虫和隐孢子虫,并用免疫荧光染色剂染色计数。结果贾第鞭毛虫的平均加标回收率为36.58%,分步加标回收率试验中“富集浓缩”这一步加标回收率最低,为57.04%;隐孢子虫的平均加标回收率为19.58%,分步加标回收率试验中“富集浓缩”这一步的加标回收率最低,为26.55%;二者的平均加标回收率均符合我国GB/T 5750—2006《生活饮用水标准检验方法》中回收率高于10%的要求。结论该方法成本低,操作相对简便,适用于普通实验室对水源水中贾第鞭毛虫和隐孢子虫的检测,其中,“富集浓缩”步骤为影响回收率的关键步骤。
OBJECTIVE To establish a method for the detection of Giardia and Cryptosporidium parvum in drinking water by means of membrane lysis-immunomagnetic separation-fluorescence staining. Methods The water samples were filtered through a mixed cellulose acetate membrane, dissolved in acetone, centrifuged and concentrated. The immunomagnetic beads were used to separate the Giardia and Cryptosporidium of the concentrated products and counted by immunofluorescence staining. Results The average recoveries of Giardia were 36.58%. The recoveries were the lowest with 57.04% in the step-by-step spike spike recovery test. The average spike recoveries of Cryptosporidium Rate of 19.58%, step-by-step spike recovery test “enrichment concentration ” this step of the spike recovery was the lowest, 26.55%; both the average spike recovery in line with our GB / T 5750-2006 “Standard Test Method for Drinking Water” in the recovery rate higher than 10% of the requirements. Conclusion The method is low cost and relatively simple to operate. It is suitable for the detection of Giardia and Cryptosporidium in drinking water in a common laboratory. Among them, the “enrichment and concentration” step is the key step affecting the recovery rate.