基于刃天青的电化学方波伏安法快速鉴别大肠杆菌的耐药性
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(1. 林产可食资源安全与加工利用湖南省重点实验室,湖南 长沙 410004;2. 中南林业科技大学食品科学与工程学院,湖南 长沙 410004;3. 九江市农产品质量安全中心,江西 九江 332000)

作者简介:

戚楚楚,女,中南林业科技大学在读硕士研究生。

通讯作者:

卢礼生(1971—),男,九江市农产品质量安全中心高级农艺师。E-mail: jjncpjczx@163.com任佳丽(1977—),女,中南林业科技大学教授,博士生导师,博士。E-mail: T20091483@csuft.edu.cn

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基金项目:

湖南省创新平台与人才计划项目(编号:2019TP1029,2021RC4032);湖南省重点研发计划项目(编号:2022SK2100)


Rapid identification of drug resistance of Escherichia coli based on electrochemical square wave voltammetry of resazurin
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(1. Hunan Key Laboratory of Forestry Edible Sources Safety and Processing, Changsha, Hunan 410004, China; 2. College of Food Science and Engineering, Central South University of Forestry and Technology, Changsha, Hunan 410004, China; 3. Jiujiang Agricultural Products Quality and Safety Monitoring Center, Jiujiang, Jiangxi 332000, China)

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    摘要:

    目的:评价5株食源性大肠杆菌临床分离株对环丙沙星、庆大霉素和氨苄西林的抗生素耐药性。方法:基于细菌能量代谢过程中的电子传递理论,引入刃天青作为氧化还原探针,建立了一种基于刃天青介导的大肠杆菌耐药性快速鉴别的电化学方波伏安法(square wave voltammetry,SWV)。结果:在1.69×102~1.69×108 CFU/mL范围内,大肠杆菌浓度与刃天青氧化峰电流(I)之间存在较好的线性关系,回归方程为Y=-0.156 0X-2.965 5,R2=0.977 5;在进行大肠杆菌耐药性测定时,参考美国临床和实验室标准协会制定的药敏试验标准,以刃天青的氧化峰电流(I0)和大肠杆菌直接与刃天青作用后得到的氧化峰电流(I-a)作为对照,测定大肠杆菌用抗生素处理后与刃天青作用得到氧化峰电流(I+a),引入细菌活性值VETA作为电化学方波伏安法鉴别细菌耐药性的指标,建立了刃天青介导的大肠杆菌电化学快速药敏试验方法;运用建立的方法检测了5种食源性大肠杆菌临床分离株对3种抗生素的耐药性,以VETA=50作为判断细菌敏感和耐药的临界值,均得到与纸片扩散法结果相符的药敏试验结果,而纸片扩散法报告结果需18 h,该法仅需2 h。结论:建立的基于刃天青介导的大肠杆菌耐药性快速鉴别的电化学方波伏安法能够准确且快速检测5株食源性大肠杆菌临床分离株对环丙沙星、庆大霉素和氨苄西林的抗生素耐药性。

    Abstract:

    Objective: To evaluate the antibiotic resistance of five foodborne Escherichia coli clinical isolates to ciprofloxacin, gentamicin and ampicillin. Methods: Based on the theory of electron transfer during bacterial energy metabolism and the introduction of resazurin as a redox probe, an electrochemical square wave voltammetry (SWV) method based on bladed tensin-mediated rapid identification of E. coli drug resistance was established. Results: The electrochemical square wave voltammetry values of E. coli at different concentrations mediated by resazurin were determined, and there was a good linear relationship between the E. coli concentration and the oxidized peak current (I) of resazurin in the range of 1.69×102~1.69×108 CFU/mL, with the regression equation of Y=-0.156 0X-2.965 5, and R2=0.977 5; When performing the determination of E. coli drug resistance, the drug sensitivity test standard was developed by Clinical and Laboratory Standards Institute (CLSI). The oxidized peak current (I0) of resazurin and the oxidized peak current (I-a) obtained by E. coli after directly interacting with resazurin were used as the control, and the oxidized peak current (I+a) obtained by E. coli after treating it with antibiotics and interacting with resazurin was determined. The value of bacterial activity VETA was introduced as VETA as an indicator for the identification of bacterial resistance by electrochemical square wave voltammetry, and established a rapid electrochemical drug sensitivity test method for E. coli mediated by resazurin. The established method was used to detect the resistance of five clinical isolates of foodborne E. coli to three antibiotics, and VETA=50 was used as the critical value for determining bacterial sensitivity and resistance, and the results of the drug sensitivity tests were all in accordance with the results of the paper diffusion method. The VETA=50 was used as the critical value for determining bacterial susceptibility and resistance, and all the results were consistent with the results of the paper diffusion method, and the paper diffusion method required 18 h to report the results, whereas this method required only 2 h. Conclusion: Our electrochemical square-wave voltammetry method based on the resazurin-mediated rapid discrimination of E. coli drug resistance is able to accurately and rapidly detect the antibiotics resistance of the five foodborne clinical isolates of E. coli to ciprofloxacin, gentamicin, and ampicillin.

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戚楚楚,卢礼生,任佳丽,等.基于刃天青的电化学方波伏安法快速鉴别大肠杆菌的耐药性[J].食品与机械,2024,40(4):47-53.
QI Chuchu, LU Lisheng, REN Jiali, et al. Rapid identification of drug resistance of Escherichia coli based on electrochemical square wave voltammetry of resazurin[J]. Food & Machinery,2024,40(4):47-53.

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  • 收稿日期:2024-01-25
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  • 在线发布日期: 2024-05-21
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