A double staining method using calcofluor white and acridine orange to differentiate life stages of Enterocytozoon hepatopenaei (EHP) on hepatopancreatic sections

文献类型： 外文期刊

第一作者： Gao, Wen

作者： Gao, Wen;Guo, Xiao-Meng;Wang, Hai-Liang;Zhao, Ruo-Heng;Xie, Guo-Si;Li, Chen;Huang, Jie;Gao, Wen;Huang, Jie;Guo, Xiao-Meng

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关键词： Enterocytozoon hepatopenaei; Calcofluor white stain; Acridine orange; Sporophyte; Plasmodium

期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）

ISSN： 0044-8486

年卷期： 2020 年 528 卷

页码：

收录情况： SCI

摘要： A double fluorescent staining method was used to differentiate life stages of Enterocytozoon hepatopenaei (EHP) with a chitin-binding dye, calcofluor white (CFW) and a nucleic acid dye, acridine orange (AO). The unreleased spores showed a significantly blue fluorescent chitin wall with weak orange-red RNA staining, while the plasmodia appeared grape cluster-like orange-red nucleic acid-containing particles without or with very weak blue fluorescence and sporophytes showed grape cluster-like orange-red particles with varying degree of blue fluorescence. The analysis of the red, blue and green intensity of different targets revealed that the ratio of relative concentration of CFW and AO (RRC/A) of plasmodia (0.36 +/- 0.15) (n = 60), sporophytes (1.25 +/- 0.41) (n = 157), and unreleased spores (3.06 +/- 1.09) (n = 21) had extremely significant differences (P < .01), which indicated an incremental relative amount of chitin to RNA in the three stages. The quantitative analysis proposed in this study indicated that double staining with CFW and AO can provide a method for differentiating various life stages of EHP in the infected hepatopancreatic cells.

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