观察改良的药物敏感性检测法对检出耐药结核分枝杆菌的影响

摘要/Abstract

摘要： 目的利用经改良的最低抑菌浓度法（IMIC）检测结核分枝杆菌临床分离株对异烟肼、利福平、乙胺丁醇、链霉素和氧氟沙星的药物敏感性，观察其对耐药结核分枝杆菌检出的影响。方法利用改良的最低抑菌浓度法检测135株临床分离株对INH、RFP、EMB、SM及OFLX的敏感性。结果在相同的药物浓度中,与接种1×10⁵/ml菌量的试验管相比135株接种1×10⁷/ml活菌数的试管中有68.89%的结果与之相同，有31.1%的结果与之不相同。判断结果由阴性转阳性的标本共有20株，（其中耐RFP的1株、耐INH的3株、耐OFLX的3株、耐EMB的5株以及耐SM的8株）约占135株受试菌株总数的14.8%。结论利用改良的最低抑菌浓度法可发现潜在的耐药结核病患者，提高耐药菌的检出率。

关键词: 分枝杆菌, 结核, 微生物敏感性试验

Abstract: Objective To develop an improved minimum inhibitory concentration (IMIC) test to detect drug-resistantMycobacterium tuberculosisclinical isolates.  Methods 135 clinical isolates detected the drug susceptibility using traditional proportional method were retested and verified their sensitivity to isoniazid (INH), rifampin (RFP), ethambutol (EMB), streptomycin (SM) and ofloxacin (OFLX) by IMIC test.  Results In the tubes with same drug concentration, 68.89% of 135 clinical isolates with 1×10⁷/ml bacterium number inoculated got the same Results as those with 1×10⁵/ml bacterium number inoculated, and 31.1% was inconsistent. 14.8% (20/135) of the isolates (including 1 RFP-resistant, 3 INH-resistant, 3 OFLX-resistant，5 EMB-resistant and 8 SM-resistant strains) were judged the Results from negative to positive. Conclusion s The improved minimum inhibitory concentration test could increase positive detectable rate of drug-resistantMycobacterium tuberculosis , and find more potential drug-resistant TB patients.

Key words: Mycobacterium tuberculos, microbial sensitivity tests

中图分类号:

R446.5

张健源, 赵雁林, 孙照刚, 姜广路, 李妍, 王甦民, 刘毅, 聂理会高铁杰, 张治国, 任卫聪, 李传友. 观察改良的药物敏感性检测法对检出耐药结核分枝杆菌的影响[J]. 中国防痨杂志, 2010, 32(6): 16-20.

Zhang Jianyuan,Zhao Yanlin,Sun Zhaogang,Jiang Guanglu,Li Yan,Wang Sumin,Liu Yi,Nie Lihui,Gao Tiejie,Zhang Zhiguo,Ren Weicong,Li Chuanyou. effect of the improved minimum inhibitory concentration test on detection of drug-resistantMycobacterium tuberculo[J]. Chinese Journal of Antituberculosis, 2010, 32(6): 16-20.

参考文献

［1］Sandgren A, Strong M, Muthukrishnan P, Weiner BK, Church GM, Murray MB. Tuberculosis Drug Resistance Mutation Database ［J］. PLoS Medicine, 2009,6(2): e1000002.
［2］ Resch SC, Salomon JA, Murray M, Weinstein MC. Cost-Effectiveness ofTreating Multidrug-Resistant Tuberculosis ［J］. PLoS Medicine, 2006,3(7): e241.
［3］ Migliori GB, Matteelli A, Cirillo D, Pai M. Diagnosis of multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis: Current standards and challenges ［J］. The Canadian Journal of Infectious Diseases & Medical Microbiology, 2008, 19(2): 169-172.
［4］ Kumanan Wilson, Michael Gardam, Christopher McDougall, Amir Attaran, and Ross Upshur WHO’s Response to Global Public Health Threats: XDR-TB ［J］. PLoS Medicine, 2007,4(7): e246.
［5］阚冠卿,张立兴,屠德华,蒋建英．积极开展结核菌原发耐药性趋势的连续观察［J］．中国防痨杂志,1994,16（1）：1-5.
［6］中国防痨协会基础专业委员会.结核病诊断实验室检验规程［M］.北京,2006.
［7］张健源,王甦民,李传友,田苗,张旭霞,赵冰,李卫民,赵雁林,邱云青,马玙.单链构象多态性与核酸杂交联合检测耐氧氟沙星的结核分枝杆菌［J］.中华结核和呼吸杂志, 2005,28(12）：841-844.
［8］ Hoek KG, Gey van Pittius NC, Moolman-Smook H, Carelse-Tofa K, Jordaan A, van der Spuy GD, Streicher E, Victor TC, van Helden PD, Warren RM. Fluorometric Assay for Testing Rifampin Susceptibility ofMycobacterium tuberculosisComplex［J］. Journal of Clinical Microbiology, 2008,46(4): 1369-1373.
［9］ Tomotada Iwamoto, Shiomi Yoshida, Katsuhiro Suzuki, Takayukj. Wada Population Structure Analysis of theMycobacterium tuberculosisBeijing Family Indicates an Association between Certain Sublineages and Multidrug Resistance［J］. Antimicrobial Agents and Chemotherapy, 2008, 52(10): 3805-3809.
［10］Ellen M,Zager,Ruth McNerney.Multidrug-resistant tuberculosis ［J］. BMC Infectious Diseases, 2008, 25:8-10.
［11］ Mariana Piuri, William R. Jacobs Jr., Graham F.Hatfull Fluoromycobacteriophages for Rapid, Specific. Sensitive Antibiotic Susceptibility Testing ofMycobacterium tuberculosis［J］.PLoS ONE,2009,4(3): e4870.
［12］ Ming-Chih Yu, Mei-Hua Wu, and Ruwen Jou Extensively Drug-Resistant Tuberculosis, Taiwan ［J］. Emerging Infectious Diseases, 2008,14(5): 849-850.
［13］ Nicole Parrish, Gerard Osterhout, Kim Dionne, Amy Sweeney, Nicole Kwiatkowski, Karen Carroll, Kenneth C. Jost Jr., James DickRapid.Standardized Method for Determination ofMycobacterium tuberculosisDrug Susceptibility by Use of Mycolic Acid Analysis［J］. Journal of Clinical Microbiology, 2007,45(12): 3915-3920.
［14］ Xinxin Yang, Eugenie Dubnau, Issar Smith, and Nicole S. Sampson Rv1106c fromMycobacterium tuberculosisis a 3β-hydroxysteroid dehydrogenase［J］. Biochemistry, 2007, 46(31): 9058-9067.
［15］ Jean-Emmanuel,Hugonnet, John S. Blanchard Irreversible Inhibition of theMycobacterium tuberculosisβ-lactamase by Clavulanate Published in final edited form as: ［J］. Biochemistry, 2007,46(43): 11998-12004.
［16］ David W. Dowdy, Richard E. Chaisson, Gary Maartens, Elizabeth L. Corbett,Susan E. Dorman Impact of enhanced tuberculosis diagnosis in South Africa: A mathematical model of expanded culture and drug susceptibility testing ［J］. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(32): 11293-11298.
［17］ Sonya S. Shin, Martin Yagui, Luis Ascencios, Gloria Yale, Carmen Suarez, Neyda Quispe,Cesar Bonilla, Joaquin Blaya, Allison Taylor, Carmen Contreras, Peter Cegielski.Scale-up of Multidrug-Resistant Tuberculosis Laboratory Services［J］. Emerging Infectious Diseases, 2008,14(5): 701-708.
［18］ Fernanda Ely, José ES Nunes, Evelyn K Schroeder, Jeverson Frazzon, Mário S Palma, Diógenes S Santos, Luiz A Basso.TheMycobacterium tuberculosisRv2540c DNA sequence encodes a bifunctional chorismate synthase［J］. BMC Biochemistry, 2008,29(9):13.
［19］ Parissa Farnia, Mohammad Reza Masjedi, Foroozan Mohammadi, Payam Tabarsei, Poopak Farnia, Ali Reza Mohammadzadeh, Parvaneh Baghei, Mohammad Varahram, Sven Hoffner, Ali Akbar Velayati.Colorimetric Detection of Multidrug-Resistant or Extensively Drug-Resistant Tuberculosis by Use of Malachite Green Indicator Dye ［J］. Journal of Clinical Microbiology,2008:796-799.
［20］ Young DB, Perkins MD, Duncan K, Barry CE.3rd Confronting the scientific obstacles to global control of tuberculosis ［J］. European Journal of Clinical Investigation, 2008,118(4): 1255-1265.

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