非可培养状态霍乱弧菌的间接免疫荧光检测

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。

Detection of Vibrio cholerae in nonculturable state by indirect immunofluorescent assay

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.