异硫氰酸荧光素-滤膜法染色贾第鞭毛虫和隐孢子虫效果比较

Study on FITC-membrane filter method of Giardia and Cryptosporidium

目的探讨贾第鞭毛虫和隐孢子虫染色方法异硫氰酸荧光素(FITC)-滤膜法,并将其与现行的国家标准方法FITC法进行比较。方法用FITC-滤膜法和FITC法分别染色贾第鞭毛虫和隐孢子虫标准体,对其染色所需时间、染色效果、检出率等逐一进行对比。结果 FITC-滤膜法的样品固定时间为5 min,远远少于FITC法的60 min,FITC-滤膜法的综合染色时间为42 min,也远少于FITC法的97 min。FITC-滤膜法对贾第鞭毛虫和隐孢子虫的检出率分别为(96.33±1.08)%和(96.67±1.70)%,高于FITC法的(94.00±2.16)%和(92.67±2.05)%,FITC-滤膜法染色可清晰看到贾第鞭毛虫和隐孢子虫的细胞核,优于FITC法。结论 FITC-滤膜法缩短了染色时间,且便于之后的分子生物学分析,可作为贾第鞭毛虫和隐孢子虫染色的替代方法使用。

Objective To explore a staining method for Giardia and Cryptosporidium, which is FITC - membrane filter method, and compared with the currently national standard method - FITC. Methods The Giardia and Cryptosporidium standard body were stained by FITC and FITC - membrane filter technique, respectively. Then to compare the staining time, staining results, and detection rates one by one. Results The fixation time of FITC - membrane filter method was 5 rain, and far less than the FITC method＇s 60 min. The comprehensive staining time of FITC -membrane filter method was 42 min, and far less than the FITC method＇s 97 rain. And the detection rates of Giardia and Cryptosporidium in FITC - membrane filter method were （96.33 ± 1.08） % and （96.67 ± 1.70） % , and higher than the FITC method＇s （94.00± 2.16） % and （92.67 ± 2, 05 ） %. The FITC - membrane filter method can clearly see the nucleus of Giardia and Cryptosporidium than the FITC method. Conclu- sion FITC - membrane filter method greatly shortens the staining time and facilitates the subsequent analysis of molecular biol- ogy. FITC - membrane filter method can be used as the basic staining method of Giardia and Cryptosporidium.