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1、湖南大学 硕士学位论文 压电生物传感仪在抗生素药敏试验中的应用研究 姓名：周娟 申请学位级别：硕士 专业：分析化学 指导教师：何凤姣 20070628 硕士学位论文 II 摘 要 抗生素是临床治疗中应用最多的一类药物。不合理地以及过量地使用抗生 素，不仅会导致耐药菌株的产生，而且会造成经济浪费，同时还会产生许多副作 用。因此，近年来抗生素的合理使用已经引起了广泛的关注。抗生素药敏试验是 一种改善抗生素使用的有效方法。 压电生物传感器因具有灵敏度高、响应谱广、操作简单、成本低廉等优点而 被广泛应用。本文充分利用串联式压电传感器对溶液电导的响应，并和微生物新 陈代谢的特点相结合，将多通道串联式压电
2、传感器应用于抗生素药敏试验的研 究， 从而拓宽了压电传感器在临床微生物中的应用。 本论文开展的研究工作如下： (1) 首次提出用多通道串联式压电传感器（MSPQC）进行抗生素药敏试验的 新方法。运用该方法测定了三种抗生素（即氨苄青霉素、头孢噻肟和庆大霉素） 对临床大肠杆菌的最低抑菌浓度， 并将临床大肠杆菌对这三种药物的敏感性进行 快速划分。MSPQC 法的试验结果与琼脂稀释法、纸片扩散法的结果基本一致。 与传统的方法相比，该方法准确、快速、操作简单，可以实时监测。由于 MSPQC 法不仅能提供最低抑菌浓度，还能快速进行敏感性划分，所以该方法有望在临床 微生物实验室得到广泛应用。 (2) 提出应
3、用多通道串联式压电传感器研究抗生素给药方案的新方法。分别 选用头孢噻肟和庆大霉素作为时间依耐性和浓度依耐性抗生素的代表药物， 研究 和讨论了不同给药剂量、给药间隔时间和给药次数对标准大肠杆菌生长的影响。 通过比较频移曲线，提出利用 FDT 评价各种给药方案的抑菌效果。这些信息将 有助于设计合理的给药方案。 (3) 提出应用多通道串联式压电传感器研究头孢噻肟和庆大霉素在体外对标 准大肠杆菌的联合作用的新方法。应用该方法进行了协同作用研究。同时还讨论 了联合用药时给药方式（同步/间隔用药、给药顺序以及给药间隔时间）对抑菌 作用的影响。研究结果将有助于制定合理的联合用药方案。 (4) 提出应用多通道
4、串联式压电传感器进行血培养吸附剂的初步筛选。运用 该法考察了三种树脂(XDA-16，CD-180 和 H-103)，以及抗凝剂 SPS 对氨苄青霉 素、头孢噻肟以及庆大霉素的吸附能力。通过比较频移曲线上的 FDT 评价吸附 抗生素的效果，并初步确定血培养吸附剂的组成。 关键词：抗生素药敏试验；大肠杆菌；氨苄青霉素；头孢噻肟；庆大霉素；多通 道串联式压电传感器 压电生物传感仪在抗生素药敏试验中的应用研究 III Abstract Antibiotics are one of the most frequently used drugs in clinical therapy. Inappropr
5、iate and overuse of antibiotics lead to not only the emergence of resistant bacteria but also economic loss and adverse reactions. So the rational use of antibiotic has aroused much attention in recent years. Antimicrobial susceptibility testing is an effective means of improving antibiotic use. Pie
6、zoelectric biosensor has been applied in many fields for its high sensitivity, broad sensing spectrum, simple operation and low cost. In this thesis, multi-channel series piezoelectric quartz crystal (MSPQC) was applied to do antimicrobial susceptibility testing, based on the response of SPQC to the
7、 change of electric parameters of medium, which related to some characteristics of the bacterial metabolism. The new methods broaden their application in clinical microbiology. The main content of this thesis is as follow: (1) A new antimicrobial susceptibility testing method by multi-channel series
8、 piezoelectric quartz crystal (MSPQC) was proposed. This method was used to test susceptibility of clinical Escherichia coli isolates against ampicillin, cefotaxime and gentamicin. Both the minimum inhibitory concentrations (MICs) and interpretive categorization of clinical E. coli isolates were det
9、ermined by the proposed method. Comparing tests were run at the same time by the agar dilution method and the disk diffusion method. The experimental results showed that MSPQC method had a good agreement with the reference methods. Compared with those methods, the MSPQC method is simple, rapid, and
10、convenient to perform. It can offer both a minimum inhibitory concentration (MIC) and an interpretive category result. It could be popularized in clinical microbiology laboratories. (2) A new sensor method based on multi-channel series piezoelectric quartz crystal (MSPQC) was proposed for studying t
11、he antibiotic dosing regimens. Cefotaxime and gentamicin were selected as time-dependence and concentration-dependence antibiotics individually. The influence of dosage, dosing time interval and dosing frequency on the growth of Escherichia coli ATCC 25922 were investigated in detail. The frequency
12、shift curves under different regimens were obtained and compared with each other. Frequency detection time （FDT） was gained to assess the inhibitory effect. The optimal dosing regimens could be made according to the obtained information. 硕士学位论文 IV (3) MSPQC method was proposed to assess the effect o
13、f gentamicin and cefotaxime combination against Escherichia coli ATCC 25922 in vitro. Synergism study was performed by the proposed method. Different combination regimens were also studied by MSPQC technique. Simultaneous/ nonsimultaneous administration, order and time interval of administration reg
14、imens were compared. The results of the present study can provide useful information to physicians in the selection of appropriate administration regimens. (4) MSPQC method was proposed to determine the antimicrobial adsorbent system. The effectiveness of three resins (XDA-16, CD-16 and H-103) and a
15、n anticoagulant SPS, to adsorb antibiotics (ampicillin, cefotaxime and gentamicin) were studied by the proposed method. Antibiotic removal was evaluated on frequency detection time (FDT). And the antimicrobial adsorbent system was determined based on the obtained results. Keywords: Antimicrobial sus
16、ceptibility testing (AST); Escherichia coli; Ampicillin; Cefotaxime; Gentamicin; Multi-channel series piezoelectric quartz crystal（MSPQC） I 湖 南 大 学 学位论文原创性声明 本人郑重声明：所呈交的论文是本人在导师的指导下独立进行研究所取 得的研究成果。除了文中特别加以标注引用的内容外，本论文不包含任何其 他个人或集体已经发表或撰写的成果作品。对本文的研究做出重要贡献的个 人和集体，均已在文中以明确方式标明。本人完全意识到本声明的法律后果 由本人承担。 作者签名： 日期： 年 月 日 学位论文版权使用授权书 本学位论文作者完全了解学校有关保留、使用学位论文的规定，同意学 校保留并向国家有关部门或机构送交论文的复印件和电子版，允许论文被查 阅和借阅。本人授权湖南大学可以将本学位论文的全部或部分内容编入有关 数据库进行检索，可以采用影印、缩印或扫描等复制手段保存和汇编本学位 论文。 本学位论文属于 1、保密，在_年解密
