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[快速生物发光抗生素敏感性测定]

[Rapid bioluminescent antibiotic susceptibility assay].

作者信息

Frundzhian V G, Ugarova N N, Blatun L A, Terekhova R P, Rusanova E V

出版信息

Antibiot Khimioter. 2009;54(7-8):8-12.

Abstract

Rapid testing of pathogen susceptibility to antibiotics is of great practical value for rational chemotherapy of pyoinflammatory deseases and postoperative complications of microbial etiology. The standard microbiological methods, i.e., the disk diffusion method and the method of serial dilutions are labour- and time-consuming (not less than 18-36 hours). The method of the authors is based on measuring bioluminescence resulting from interaction of adenosine-5'-triphosphate (ATP) and ATP reagent, a standard reaction mixture of firefly luciferase (an enzyme) and luciferin. The bioluminescence intensity is proportional to the ATP concentration in the reaction mixture and the ATP concentration is proportional to the number of the pathogen viable cells in the sample. The bioluminescence intensity value in the pathogen suspension aliquots with and without (control) the antibiotic were compared after the incubation for 5 hours and the coefficient of the microbial cell growth inhibition was calculated. Satisfactory correlation (R2 > 88%) of the results of the bioluminescent assay and the assay with the disk diffusion method and the method of serial dilutions was observed.

摘要

快速检测病原体对抗生素的敏感性对于脓性炎症疾病和微生物病因引起的术后并发症的合理化疗具有重要的实际价值。标准的微生物学方法,即纸片扩散法和连续稀释法,既耗时又费力(不少于18 - 36小时)。作者的方法基于测量由腺苷 - 5'-三磷酸(ATP)与ATP试剂相互作用产生的生物发光,ATP试剂是萤火虫荧光素酶(一种酶)和荧光素的标准反应混合物。生物发光强度与反应混合物中的ATP浓度成正比,而ATP浓度与样品中病原体活细胞的数量成正比。在孵育5小时后,比较添加和不添加(对照)抗生素的病原体悬浮液等分试样中的生物发光强度值,并计算微生物细胞生长抑制系数。观察到生物发光测定结果与纸片扩散法和连续稀释法测定结果具有良好的相关性(R2 > 88%)。

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