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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。
Objective To establish an indirect immunofluorescence assay for detecting Vibrio cholerae (O1 population, O139 population) that are not cultured in water. Methods Antigen fragments were prepared from the bacteria culture of Vibrio cholerae that were artificially transformed in this laboratory. The mixed diagnostic sera (anti-O1 group, O139 group) were used as primary antibody and fluorescein isothiocyanate (FITC) labeled pig Anti-rabbit IgG as secondary antibody, indirect immunofluorescence assay. Results The method could successfully detect Vibrio cholera in non-culturable water with the detection limit of about 104 cells / 400 ml. Conclusion The method has good specificity and sensitivity and can be used as a detection method of Vibrio cholerae in non-culturable state in natural water for the ecological research of Vibrio cholerae.