A flow cytometry method for detecting multi-vibrios using anti-OmpW monoclonal antibody

Abstract:

[Objective] To establish the flow cytometry (FCM) method for detecting multi-vibrio based on anti-OmpW monoclonal antibody. [Methods] With the rate of cell stainning as an indicator, optimize the reactived conditions of anti-OmpW mAb concentration and reaction time for detecting Vibrio parahaemolyticus by flow cytometry. Evaluate the accuracy, detection limits and precision of flow cytometry by comparing the munber of bacteria calculated by FCM with that in the culture. Based on the obove flow cytometry method, analysize and identify the mAb specificity to the five vibrios. [Results] The optimized mAb concentraion was 20 mg/L, and the optimized reaction time of mAb was 60min when Vibrio parahaemolyticus was detected by flow cytometry. The FCM method could provide high reliability which could specifically recognize the five vibrios in the appropriate cell concentration of 104?107cells/mL. To detect different cell concentration samples repeatedly, the coefficient of variation was less than 7%. [Conclusion] The established flow cytometry method based on anti-OmpW mAb could quickly and accurately detect multiple vibrios.

Key words:Monoclonal antibody, multi-Vibrio, flow cytometry