通过定量实时聚合酶链反应测定铜绿假单胞菌多药外排泵
Measurement of Pseudomonas aeruginosa multidrug efflux pumps by quantitative real-time polymerase chain reaction.
作者信息
Yoneda Kazuhiko, Chikumi Hiroki, Murata Takeshi, Gotoh Naomasa, Yamamoto Hiroyuki, Fujiwara Hiromitsu, Nishino Takeshi, Shimizu Eiji
机构信息
Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Faculty of Medicine, Tottori University, 36-1 Nishi-machi, Yonago-shi, Tottori-ken 683-0805, Japan.
出版信息
FEMS Microbiol Lett. 2005 Feb 1;243(1):125-31. doi: 10.1016/j.femsle.2004.11.048.
Multidrug efflux pumps contribute to multiple antibiotic resistance in Pseudomonas aeruginosa. Pump expression usually has been quantified by Western blotting. Quantitative real-time polymerase chain reaction has been developed to measure mRNA expression for genes of interest. Whether this method correlates with pump protein quantities is unclear. We devised a real-time PCR for mRNA expression of MexAB-OprM and MexXY-OprM multidrug efflux pumps. In laboratory strains differing in MexB and MexY expression and in several clinical isolates, protein and mRNA expression correlated well. Quantitative real-time PCR should be a useful alternative in quantitating expression of multidrug efflux pumps by P. aeruginosa isolates in clinical laboratories.
多药外排泵导致铜绿假单胞菌产生多重抗生素耐药性。泵的表达通常通过蛋白质免疫印迹法进行定量。定量实时聚合酶链反应已被开发用于测量感兴趣基因的mRNA表达。该方法是否与泵蛋白量相关尚不清楚。我们设计了一种用于检测MexAB - OprM和MexXY - OprM多药外排泵mRNA表达的实时PCR方法。在MexB和MexY表达不同的实验室菌株以及几种临床分离株中，蛋白质和mRNA表达相关性良好。定量实时PCR应该是临床实验室中对铜绿假单胞菌分离株的多药外排泵表达进行定量的一种有用替代方法。
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