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[BD Phoenix100系统与多黏菌素肉汤纸片洗脱法对革兰阴性菌进行多黏菌素药敏试验的评估]

[Evaluation of the BD Phoenix100 System and Colistin Broth Disk Elution Method for Antimicrobial Susceptibility Testing of Colistin Against Gram-negative Bacteria].

作者信息

Koyuncu Özyurt Özlem, Özhak Betil, Öğünç Dilara, Yıldız Emre, Çolak Dilek, Günseren Filiz, Öngüt Gözde

机构信息

Akdeniz University Faculty of Medicine, Department of Medical Microbiology, Antalya, Turkey.

Akdeniz University Faculty of Medicine, Department of Infectious Diseases and Clinical Microbiology, Antalya, Turkey.

出版信息

Mikrobiyol Bul. 2019 Jul;53(3):254-261. doi: 10.5578/mb.68066.

Abstract

Infections with multidrug resistant gram-negative bacteria is a growing problem especially in health care settings. Colistin is one of the last resort antibiotics for such infections in which treatment options are limited. Increasing resistance to colistin is a global problem. Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) study groups have recommended the ISO-standard broth microdilution method (20776-1) as the reference method for the determination of colistin susceptibility. Since the broth microdilution method is not a practical method, it is rarely used in routine clinical microbiology laboratories, yet simple and accurate phenotypic detection methods for the determination of colistin resistance in routine microbiology laboratories are not precisely defined. The aim of this study was to evaluate BD Phoenix100 (Becton Dickinson, USA) system and colistin broth disk elution method for the detection of in vitro activity of colistin against gram-negative bacteria. A total of 419 gram-negative bacteria, including 199 Klebsiella pneumoniae, 163 Acinetobacter baumannii, 34 Escherichia coli, 20 Enterobacter spp., and three Citrobacter spp. isolates which were isolated from various clinical samples in our hospital between 2016-2018 were tested. The broth microdilution method was used as the reference method applying ISO-standard broth microdilution methods (20776-1) and CLSI/EUCAST recommendations. For colistin broth disk elution method, final concentrations of 0 (growth control), 1, 2 and 4 μg/ml were obtained by adding 10 μg colistin disks to four tubes containing 10 ml cation-adjusted Mueller Hinton broth per isolate. After incubation at room temperature for 30 minutes, 50 μl of standardized inoculum suspensions were added to the tubes. Colistin minimum inhibitor concentration (MIC) values were read visually after 16-20 hours of incubation at 35°C in ambient air. Manufacturer's recommendations were followed for BD Phoenix100 system. The categorical agreement between the reference broth microdilution method and the colistin broth disk elution method was 99.3%, very major error and major error rates were 0.2% and 0.5%, respectively. For BD Phoenix100 system, the categorical agreement was 95%, with a very major error rate of 5%. Our results showed that colistin broth disc elution method worked well compared to the reference broth microdilution method. The BD Phoenix100 system, with a high very major error rate, does not reliably distinguish colistin-resistant and colistin-susceptible strains.

摘要

多重耐药革兰氏阴性菌感染是一个日益严重的问题,尤其是在医疗环境中。对于治疗选择有限的此类感染,黏菌素是最后的抗生素之一。对黏菌素耐药性的增加是一个全球性问题。临床和实验室标准协会(CLSI)以及欧洲抗菌药物敏感性试验委员会(EUCAST)研究小组推荐采用ISO标准肉汤微量稀释法(20776-1)作为测定黏菌素敏感性的参考方法。由于肉汤微量稀释法并非实用方法,在常规临床微生物实验室中很少使用,然而,常规微生物实验室中用于测定黏菌素耐药性的简单且准确的表型检测方法尚未明确界定。本研究的目的是评估BD Phoenix100(美国BD公司)系统和黏菌素肉汤纸片洗脱法检测黏菌素对革兰氏阴性菌的体外活性。对2016年至2018年期间从我院各种临床样本中分离出的总共419株革兰氏阴性菌进行了检测,其中包括199株肺炎克雷伯菌、163株鲍曼不动杆菌、34株大肠埃希菌、20株肠杆菌属菌株和3株柠檬酸杆菌属菌株。采用ISO标准肉汤微量稀释法(20776-1)并遵循CLSI/EUCAST建议,将肉汤微量稀释法用作参考方法。对于黏菌素肉汤纸片洗脱法,通过向每株菌的四个装有10 ml阳离子调节的Mueller Hinton肉汤的试管中加入10 μg黏菌素纸片,获得最终浓度为0(生长对照)、1、2和4 μg/ml。在室温下孵育30分钟后,向试管中加入50 μl标准化接种物悬液。在35°C环境空气中孵育16至20小时后,目视读取黏菌素最低抑菌浓度(MIC)值。BD Phoenix100系统遵循制造商的建议。参考肉汤微量稀释法与黏菌素肉汤纸片洗脱法之间的分类一致性为99.3%,极重大误差率和重大误差率分别为0.2%和0.5%。对于BD Phoenix100系统,分类一致性为95%,极重大误差率为5%。我们的结果表明,与参考肉汤微量稀释法相比,黏菌素肉汤纸片洗脱法效果良好。BD Phoenix100系统极重大误差率较高,不能可靠地区分黏菌素耐药和敏感菌株。

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