摘要
为了建立一种可以应用于水产养殖现场的药敏快速检测方法,以迟钝爱德华氏菌JF-1为试验菌株,在MHB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,优选了迟钝爱德华氏菌的增菌液,在此基础上利用阿尔玛蓝作为细菌生长指示剂,选择11种水产常见药物并设置了8个倍比稀释的质量浓度梯度,制备了迟钝爱德华氏菌快速药敏检测微孔板,板内设有药敏检测区、生长对照区及空白对照区。经显色法及分光光度法药敏检测试验结果比对,选定1×105个/mL菌液密度作为快速药敏检测的接菌密度,将该密度菌液接种于制备的快速药敏检测微孔板进行药敏检测,同时以分光光度法及试管稀释法检测结果作为比对,结果显示在接种12h后,显色法测定的所有药物对菌株JF-1的最小抑菌质量浓度与分光光度法完全吻合,但与接种24h后观察的稀释法在氨苄青霉素和复方新诺明两种药物的最小抑菌质量浓度上存在微小偏差。试验结果显示,本研究建立的迟钝爱德华氏菌快速药敏检测方法能够快速、简便、准确地对该菌进行药敏检测。
Edwardsiella tarda as an etiological agent of edwardsiellosis, a serious systematic disease, is responsible for large economic losses in aquaculture. Recently, a number of E. tarda strains isolated have exhibited resistance to multiple antibiotics, and lead to difficulty to effective clinic therapy. In order to exert effective and in-time therapeutic measures against edwardsiellosis, rapid antimicrobial susceptibility testing methods are urgently needed to be developed for scientific and reasonable selection of antibiotics in therapy. Based on MH broth, enrichment broth for E. tarda strain JF-1 was optimized by addition of fish peptone and inorganic salt ions, and Alamar Blue was employed as the color indicator, and 11 kinds of antibiotics were inoculated into the 96-well microplates at 8 different concentrations by doubling dilution for construction of rapid antibiotic sensitivity testing （AST） plate which contained AST region, growth control region and blank region. Compared with the color-developing method and spectrophotometric assay, the strain JF-1 suspension at 1 ×10^5 cells/mL as optimal inoculation concentration was inoculated into the prepared rapid AST plate, and detected by a spectrophotometric assay and tube dilution method for AST in parallel as comparison. 12 h post inoculation, the MIC data obtained by color-developing method on the rapid AST plate was in accordance with the results acquired by spectrophotometric assay, and showed a little deviation with the results obtained by tube dilution method at 24 h post inoculation. The results showed that the rapid AST method developed based on color-developing was well supported by spectrophotometric assay and tube dilution method, although there is a little deviation between the results obtained from tube dilution method and the rapid AST method. The wells in the microplate have intrinsically a large surface-to-volume （S/V） ratio resulting in a higher oxygen dissolving efficiency, which would greatly benefit the bacterial propagation and show a
出处
《水产科学》
CAS
北大核心
2014年第1期22-28,共7页
Fisheries Science
基金
山东省科技攻关计划项目(2010GHY10503)
山东省优秀中青年科学家科研奖励基金计划项目(BS2010NY021)
关键词
迟钝爱德华氏菌
药敏试验
阿尔玛蓝
显色法
快速检测
Edwardsiella tarda antibiotic sensitivity testing Alamar Blue color-developed method rapid detection
