生物膜电极的制备及其在抗菌药物有效性检测中的应用  

Preparation of Biofilm Electrode and Application in the Detection of Antimicrobial Efficacy

作　　者：马烨 何迪 陈立志 王晶晶 王超 刘畅 MA Ye;HE Di;CHEN Li-zhi;WANG Jing-jing;WANG Chao;LIU Chang(College of Pharmacy,Jinzhou Medical University,Jinzhou 121000,China)

机构地区：[1]锦州医科大学药学院,辽宁锦州121000

出　　处：《化学试剂》2020年第7期767-772,共6页Chemical Reagents

基　　金：辽宁省大学生创新创业训练项目(20151016000-0007)。

摘　　要：首先采用电聚合法制备了聚中性红修饰玻碳电极(PNR/GCE),在此基础上,以壳聚糖(CS)为成膜材料,石墨烯(GR)为导电纳米材料,通过掺杂方式将模型微生物E.coli固定于PNR/GCE上,制备生物膜电极GR/CS/E.coli/PNR/GCE。采用正交实验和方差分析考察GR浓度、CS浓度和E.coli浓度3个因素对GR/CS/E.coli/PNR/GCE性能的影响。继而借助中性红可作为电子媒介体的特性,通过考察E.coli活性与抗菌药物浓度的关系构建一种新型体外药效检测方法,用以快速检测抗菌药物有效性。GR终浓度0.50 mg/mL,CS终浓度0.75%(w/v%),E.coli终浓度0.50 g/mL为电极制备的最佳条件。将最优条件下制备的GR/CS/E.coli/PNR/GCE应用于庆大霉素、左氧氟沙星、头孢曲松钠、哌拉西林钠及万古霉素5种抗菌药物的有效性测定,其中前4种抗菌药物对E.coli的10%抑菌浓度(IC10)分别为15.81、19.62、65.74、62.47 mg/L,而针对革兰氏阳性菌的万古霉素对E.coli基本无抑制作用。所得结果与经典的纸片扩散法所得结果具有良好的相关性。Firstly,poly neutral red modified glassy carbon electrode(PNR/GCE)was prepared by electropolymerization.On this basis,chitosan(CS)as a film-forming material and grapheme(GR)as a conductive nanomaterial,biofilm electrode GR/CS/E.coli/PNR/GCE was prepared by immobilization of model microorganism E.coli to PNR/GCE by doping method.Orthogonal design and analysis of variance were used to investigate the influence of three factors,such as GR concentration,CS concentration and E.coli concentration,on the GR/CS/E.coli/PNR/GCE performance.Then a new method for rapid detection of antimicrobial efficacy in vitro was constructed where NR as an electron mediator was employed in investigating the relationship between the activity of E.coli and the concentration of antimicrobials.The optimum conditions for electrode preparation were as follows:the final concentrations of GR,CS and E.coli were 0.5 mg/mL,0.75%(w/v%),0.50 g/mL,respectively.The prepared GR/CS/E.coli/PNR/GCE under the optimal conditions was applied to detect the efficacies of gentamycin,levofloxacin,ceftriaxone sodium,piperacillin sodium and vancomycin,which antimicrobial agents mentioned above,such as gentamycin,levofloxacin,ceftriaxone sodium and piperacillin sodium,showed 10%inhibition on E.coli with the inhibiting concentration(IC10)of 15.81,19.62,65.74,62.47 mg/L,respectively.In contrast,vancomycin,as an antibiotic against gram-positive bacteria,had little inhibitory effect on E.coli cells.The results obtained above were in good correlation with the results obtained by classical disk diffusion method.

关 键 词：聚中性红 修饰电极 微生物固定化 生物膜电极 抗菌药物有效性检测 

分 类 号：O657.1[理学—分析化学]