非可培养状态霍乱弧菌的间接免疫荧光检测

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

doi:10.11847/zgggws2005-21-12-16

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

摘要

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 immunoflour escent assay(IFA)to detect viable but nonculaurable state Vibrio cholerate O1 and O 139.
Methods Viable but no nculturable 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 isothiocy anate(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 10⁴ cells/400ml.
Conclusion The IFA method was sensitive and specific for detecting Vibrio choler ae in viable but nonculturable state, and was reliable for labor atory diagnosis of nonculturable state Vibrio cholerae that exist in natural water bodies, providing a effective tool for the ecological study of Vibrio cholerae.

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