全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

用线性梯度平板准确测定药物敏感性和分离抗药性菌株

, PP. 748-755

Keywords: 线性梯度平板,E.coli,恩诺沙星,MIC,抗性菌

Full-Text   Cite this paper   Add to My Lib

Abstract:

菌群在药物的最低抑菌浓度附近的动力学过程是抗生素药理学研究的核心问题.建立一种能确定精确的MIC且又能准确分离抗药性菌株的方法,是目前临床对药敏实验新的要求.根据Fick扩散定律制备了线性梯度平板:将15mL含适当浓度恩诺沙星的琼脂培养基在9cm培养皿中倾斜凝固,刚好覆盖整个平板底面,然后水平放置,再在其上层加入同样体积的无药琼脂培养基,凝固12h后,药物浓度达到扩散平衡而呈均匀连续线性梯度.通过实测验证药物浓度在平板表面呈线性梯度分布.将待检E.coli菌群均匀涂布在梯度平板上,培养12h后,随恩诺沙星浓度提高依次形成连续密集小菌落区和离散大菌落区,根据两区域的分界线可以确定菌群自然形成的真实的MIC,与常规药敏实验方法测定结果一致.大菌落重新涂布高梯度平板,分界线显著上升,并检测出抗药性基因突变,表明该方法很容易筛选出菌群中的抗药性菌株.梯度平板可以方便地呈现整个菌群在MIC附近的动力学过程和遗传生理变化,并预警该抗生素使用后可能出现的抗药性,从而指导临床抗菌药物的选择和使用.

References

[1]  1 Wheat P F. History and development of antimicrobial susceptibility testing methodology. J Antimicrob Chemother, 2001, 48: S1–S4??
[2]  2 Bauer A W, Kirby W M, Sherris J C, et al. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol, 1966, 45: 493–496
[3]  3 CLSI. Performance standards for antimicrobial susceptibility testing. Twentieth. Informational Supplement. CLSI document M100-S20. Wayne, PA: Clinical and Laboratory Standard Institute, 2010
[4]  4 World Health Organization. Standardization of methods for conducting microbiology sensitivity tests. Second report of the expert committee on Antibiotics. WHO Technical Report Series, No. 210. WHO, Geneva, 1961
[5]  5 Gould I M. Towards a common susceptibility testing method? J Antimicrob Chemother, 2000, 45: 757–762
[6]  6 杨启文, 王辉, 徐英春, 等. 中国教学医院细菌耐药监测研究及现状介绍. 中国临床药理学杂志, 2008, 24: 570–573
[7]  7 肖永红. 卫生部全国细菌耐药监测网(Mohnarin)介绍. 中国抗生素杂志, 2008, 33: 577–588
[8]  8 Taubes G. The Bacteria fight back. Science, 2008, 18: 321: 356–361
[9]  9 Richard G W, Lewis K, Salyers A A, et al. Bacterial Resistance to Antimicrobials, 2nd ed. Boca Raton: CRC Press, 2007
[10]  10 朱小玲, 齐静, 白华, 等. 山东省动物源大肠杆菌多重抗药性及其遗传稳定性研究. 中国卫生检疫杂志, 2009, 19: 1473–1476
[11]  11 Bryson V, Szybalski W. Microbial selection. Sience, 1952, 116: 45–51??
[12]  12 Shimkets L J, Dworkin M, Keller K H. A method for establishing stable concentration gradients in agar suitable for studying chemotaxis on a solid surface. Can J Microbiol, 1979, 25: 1460–1467??
[13]  13 张金儿, 朱江萍, 刘义雄, 等. 从氯霉素抗性突变株筛选赤霉素高产菌株. 中国抗生索杂志, 2005, 30: 301–303
[14]  14 陈双雅, 邓子新. 吸水链霉菌井冈变种JG5008 转化系统的初建. 应用与环境生物学报, 2000, 6: 267–270
[15]  15 Philip Nelson, 著. 黎明, 戴陆如, 译. 生物物理学: 能量信息生命. 上海: 上海科学技术出版社, 2006. 116–118
[16]  18 Bagel S, Hullen V, Wiedemann B, et al. Impact of gyrA and parC mutations on quinolone resistance, doubling time, and supercoiling degree of Escherichia coli. Antimicrob Agents Chemother, 1999, 43: 868–875
[17]  19 刘玉庆, 张怀强, 胡明, 等. 药敏实验方法的局限性及改进的建议. 山东大学学报(医学版), 2011, 49: 124–132
[18]  20 Brown D F. Determination of MICs by the E test. J Antimicrob Chemother, 1992, 29: 455??
[19]  21 Hill G B, Schalkowsky S. Development and evaluation of the spiral gradient endpoint method for susceptibility testing of anaerobic gram-negative bacilli. Rev Infect Dis, 1990, 12: S200–S209??
[20]  22 Cooper K E. Theory of antibiotic inhibition zones in agar media. Nature, 1995, 175: 510–511
[21]  23 Finn R K. Theory of agar diffusion methods for bioassay. Anal Chem, 1959, 31: 975–977??
[22]  24 Awerbuch T E, Lustman L. A mathematical model for determining minimum inhibitory concentration (MIC) via diffusion assay. J Theory Biol, 1987, 129: 219–230??
[23]  25 Ringertz S, Kronvall G. On the theory of the disk diffusion test. APMIS, 1988, 96: 484–490??
[24]  26 Drugeon H B, Juvin M E, Caillon J, et al. Assessment of formulas for calculating critical concentration by the agar diffusion method. Antimicrob Agents Chemother, 1987, 31: 870–875
[25]  27 Anderson B. Improved susceptibility disk assay method employing an agar overlay technique. Antimicrob Agents Chemother, 1978, 14: 761–764
[26]  28 Bonev B, Hooper J, Parisot J. Principles of assessing bacterial susceptibility to antibiotics using the agar diffusion method. J Antimicrob Chemother, 2008, 61: 1295–1301??
[27]  29 Andrews J M. The development of the BASA standsrdized method of disk diffusion testing. Antimicrob Agent Chemoth, 2001, 48: 29–42??
[28]  30 Turnidge J, Paterson D L. Setting and revising antibacterial susceptibility breakpoints. Clin Microbiol Rev, 2007, 20: 391–408??
[29]  31 陈秀枢. 纸片扩散法抗菌素敏感性实验的局限性及其补充方法. 临床检验杂志, 1994, 12: 2–5
[30]  32 Jorgensen J H, Crawford S A, Fulcher L C, et al. Multilaboratory Evaluation of disk diffusion antimicrobial susceptibility testing of Neisseria meningitidis isolates. J Clinical Microbiology, 2006, 44: 1744–1754??
[31]  33 Zhao X, Drlica K. Restricting the selection of antibiotic-resistant mutants: measurement and potential uses of the mutant selection window. J Infect Dis, 2002, 185: 561–565??
[32]  34 张怀强, 刘玉庆, 刘波, 等. 分批培养条件下细菌群体生长阶段的区分及生长参数的确定. 中国科学C 辑: 生命科学, 2005, 35 502–512
[33]  35 刘玉庆, 张怀强, 沈建忠, 等. 大肠杆菌群体的生理异质性对药敏实验的影响. 中国科学C 辑: 生命科学, 2007, 37: 524–529
[34]  36 张怀强, 赵越, 何秀丽, 等. 构建大肠杆菌药敏实验新方法的探索. 中国科学: 生命科学, 2010, 40: 1063–1073
[35]  37 Cooper K E, Linton A H, Sehgal S N. The effect of inoculum size on inhibition zones in agar media using staphylococci and streptomycin. J Gen Microbiol, 1958, 18: 670–687
[36]  38 Firsov A A, Vostrov S N, Kononenko O V, et al. Prediction of the effects of inoculum size on the antimicrobial action of trovafloxacin and ciprofloxacin against Staphylococcus aureus and Escherichia coli in an in vitro dynamic model. Antimicrob Agents Chemoth, 1999, 43: 498–502
[37]  39 杨纪珂, 齐翔林, 陈霖. 生物数学概论. 北京: 科学出版社, 1982. 640–722
[38]  16 Liu Y Q, Zhang Y Z, Gao P J. Novel Concentration-Killing Curve method for estimation of bactericidal potency of antibiotics in an in vitro dynamic model. Antimicrob Agents Chemother, 2004, 48: 3884–3891??
[39]  17 王似菊, 沈向忠, 宗俭. 诺氟沙星的荧光分光光度测定法. 中国医药工业杂志, 1993, 24: 411–413

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133