摘要
目的探讨基于沙眼衣原体全菌体兔血清抗体的间接免疫荧光法检测沙眼衣原体包涵体抗原的研究。方法将6×103IFU沙眼衣原体接种于Hep-2细胞,培养72 h,PCR检测验证;以兔抗沙眼衣原体全菌体血清抗体为一抗,用免疫荧光法检测沙眼衣原体包涵体抗原,同时采用传统的Giemsa染色、Lugol′s碘染对沙眼衣原体包涵体进行鉴定,并统计和比较其包涵体的检出率。结果经PCR鉴定为阳性的沙眼衣原体感染的Hep-2细胞,间接免疫荧光检测在细胞浆中出现绿色荧光,检出率为28.84%;经Giemsa染色和Lugol′s碘染验证胞浆中存在典型的包涵体结构,检出率分别为15.19%和17.36%,与免疫荧光法比较差异有统计学意义(P<0.01)。结论基于兔抗沙眼衣原体全菌体血清的间接免疫荧光法可显著提高沙眼衣原体包涵体的检出率。
Objective To investigate use of the indirect immunofluorescence（IIF） method with rabbit anti-Chlamydia trachomatis serum antibodies to detect whole cell inclusions of C.trachomatis.Methods Hep-2 cells were inoculated with 6×103 IFU of C.trachomatis.After 72 h,the presence of C.trachomatis in Hep-2 cells was confirmed by PCR.Then,whole cell inclusions of C.trachomatis were detected by IIF with rabbit anti-C.trachomatis serum antibodies as the first antibody.At the same time,traditional Giemsa staining and Lugol＇s iodine staining were also used to identify whole cell inclusions,and the rate of detection of inclusion bodies was calculated and compared.Results With an indirect immunofluorescence assay,green fluorescence appeared in the cytoplasm of Hep-2 cells that were identified as C.trachomatis-positive by PCR,and the rate of detection of the whole cell inclusions was 28.86%.At the same time,Giemsa and Lugol＇s iodine staining revealed typical whole cell inclusions in the cytoplasm of Hep-2 cells infected with C.trachomatis.The difference between the rate of detection of indirect immunofluorescence and that of Giemsa staining（15.19%） or Lugol＇s iodine staining（17.36%） was statistically significant（P0.05）.Conclusion An indirect immunofluorescence assay based on rabbit anti-C.trachomatis serum antibodies was used to identify whole cell inclusions of C.trachomatis and significantly increased the rate of detection of C.trachomatis.
出处
《中国病原生物学杂志》
CSCD
2010年第8期561-563,F0004,共4页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.30972669)
浙江省教育厅基金资助项目(No.Y200803445)
关键词
间接免疫荧光法
沙眼衣原体
包涵体
鉴定
Indirect immunofluorescence
Chlamydia trachomatis
inclusion
identification
