建立并验证三重实时荧光定量 PCR 法检测鲍氏不动杆菌外排泵介导的抗生素耐药性。
Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei.
机构信息
Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia.
Faculty of Science, Health, Education & Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia.
出版信息
Future Microbiol. 2018 Sep;13(12):1403-1418. doi: 10.2217/fmb-2018-0155. Epub 2018 Sep 26.
Abstract
AIM
To develop a probe-based triplex quantitative real-time PCR assay to simultaneously detect the upregulation of the efflux pumps AmrAB-OprA, BpeAB-OprB and BpeEF-OprC in Burkholderia pseudomallei strains exhibiting increased minimum inhibitory concentrations toward meropenem, doxycycline or trimethoprim-sulfamethoxazole.
METHODS
The triplex assay was developed and subsequently tested on RNA isolated from eight clinical and eight laboratory-generated B. pseudomallei mutants harboring efflux pump regulator mutations.
RESULTS
The triplex assay accurately detected efflux pump upregulation in all clinical and laboratory mutants, which corresponded with decreased antibiotic susceptibility or antibiotic resistance.
CONCLUSION
Rapid detection of antibiotic resistance provides clinicians with a tool to identify potential treatment failure in near real time, enabling informed alteration of treatment during an infection and improved patient outcomes.
摘要
目的
开发一种基于探针的三重实时定量 PCR 检测方法，以同时检测具有增加的美罗培南、多西环素或复方磺胺甲噁唑最低抑菌浓度的伯克霍尔德氏菌假单胞菌菌株中流出泵 AmrAB-OprA、BpeAB-OprB 和 BpeEF-OprC 的上调。
方法
开发了三重检测方法，并随后对携带流出泵调节剂突变的 8 株临床和 8 株实验室产生的伯克霍尔德氏菌假单胞菌突变株分离的 RNA 进行了测试。
结果
三重检测方法准确地检测到所有临床和实验室突变株中流出泵的上调，这与抗生素敏感性降低或抗生素耐药性相关。
结论
快速检测抗生素耐药性为临床医生提供了一种工具，可在近实时的基础上识别潜在的治疗失败，从而在感染期间及时调整治疗，并改善患者的预后。
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