一种水产弧菌快速药敏检测方法的建立

Establishment of a Rapid Antibiotic Sensitivity Testing Method for Vibrio Species

【目的】实现弧菌药物敏感性的现场快速检测。【方法】以副溶血性弧菌、溶藻弧菌及灿烂弧菌3种弧菌为试验菌株,在Mueller-Hinton(MH)培养基基础上通过添加鱼肉蛋白胨及调节无机盐离子浓度,优选弧菌的增菌液。同时利用阿尔玛蓝作为颜色指示剂,选择11种水产常见药物并设置8个倍比稀释的浓度梯度,结合运用海藻糖作为包被保护剂,制备弧菌快速药敏检测微孔板,板内设有药敏检测区、生长对照区及质控区。【结果】利用药敏微孔板法及传统的试管稀释法分别对副溶血性弧菌、溶藻弧菌及灿烂弧菌进行药敏检测,结果显示微孔板法测定的所有药物对3种弧菌的MIC值与试管稀释法完全吻合。【结论】本研究建立的药敏微孔板法能够实现对3种弧菌药敏特性的快速、简便、准确检测,研究结果将为水产病原微生物的现场快速选药技术提供重要参考。

【Objective】In order to exert effective and in-time therapeutic measures against Vibriosis,rapid antimicrobial susceptibility testing methods were urgently needed to be developed for scientific and reasonable selection of antibiotics in therapy.【Methods】The Vibiro parahaemolyticus,V.alginolyticus and V.splendidus were selected as test strains.On the basis of Mueller-Hinton（MH） medium,enrichment broth for Vibrio species was optimized by adding fish protein peptone and adjusting the concentration of the inorganic salt ion.At the same time,for the construction of rapid antibiotic sensitivity testing（AST）plate,Alamar Blue and Trehingose were employed as the color indicator and coat stabilizer respectively,and 11 kinds of common use medicines in aquaculture were selected and 8 different concen-trations by doubling dilution were set.The rapid AST plate contained AST region,growth control region and quality control region.【Results】The MIC data of the 11 kinds of antibiotics to Vibiro parahaemolyticus,V.alginolyticus and V.splendidus obtained by the rapid AST plate method were in accordance with the results acquired by tube dilution method,which indicated that the rapid AST method developed based on color-developing was accurate and feasible for antimicrobial susceptibility testing.【Conclusion】The rapid AST plate method established in this study can realize the rapid,simple and accurate detection of the antibiotic sensitivity characteristics of three kinds of Vibriospecies,and the research results will provide an important reference for the on-site rapid medicine selection technology of aquatic pathogenic microorganisms.