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非可培养状态霍乱弧菌的间接免疫荧光检测

Detection of Vibrio cholerae in nonculturable state by indirect immunofluorescent assay

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【作者】 李孝权王鸣易鸿刘于飞莫自耀黄冰蒋力云刘远范健文李钏华邓志爱柴巧学

【Author】 LI Xiaoquan,WANG Ming,YI Hong,et al.Guangzhou Center For Disease Control and Prevention(Guangzhou 510080,China)

【机构】 广东省广州市疾病预防控制中心广州市荔湾区疾病预防控制中心遵义医学院口腔医学系 510080510080

【摘要】 目的建立检测水中活的非可培养状态霍乱弧菌(O1群、O139群)的间接免疫荧光法。方法用本实验室人工转化的非可培养状态霍乱弧菌的菌液制成抗原片,用混合诊断血清(抗O1群、O139群)作为一抗,异硫氰酸荧光素(FITC)标记猪抗兔IgG作为二抗,进行间接免疫荧光检测。结果该方法可以成功检测出水中非可培养状态霍乱弧菌,其检测下限约为104cells/400 ml。结论该方法具有较好的特异性和灵敏度,可作为自然水体中非可培养状态霍乱弧菌的检测方法,用于霍乱弧菌的生态研究。

【Abstract】 Objective To develop a diagnostic test based on indirect immunoflourescent assay(IFA) to detect viable but nonculaurable state Vibrio cholerate O1 and O139.Methods Viable but nonculturable state(VBNC) Vibrio cholerae transformed artificially in our laboratory were utilized as antigen on slide.Mixed diagnostic antisera(against serotype O1 and O139)was employed as direct antibody and fluorescein isothiocyanate(FITC)conjugated antibody to rabbit IgG as indirect antibody in IFA.Results This method could detect viable but nonculturable state Vibrio cholerae in water successfully,and the low limit of detection was 10~4 cells/400ml.Conclusion The IFA method was sensitive and specific for detecting Vibrio cholerae in viable but nonculturable state,and was reliable for laboratory diagnosis of nonculturable state Vibrio cholerae that exist in natural water bodies,providing a effective tool for the ecological study of Vibrio cholerae.

【基金】 广东省医学科研基金项目(A2002957)
  • 【文献出处】 中国公共卫生 ,China Public Health , 编辑部邮箱 ,2005年12期
  • 【分类号】R392.1
  • 【被引频次】17
  • 【下载频次】199
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