Pseudomonas aeruginosa MexAB OprM active efflux efflux phenotype screening and genetic testing.docVIP

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Pseudomonas aeruginosa MexAB OprM active efflux efflux phenotype screening and genetic testing.doc

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Pseudomonas aeruginosa MexAB OprM active efflux efflux phenotype screening and genetic testing

Pseudomonas aeruginosa MexAB OprM active efflux efflux phenotype screening and genetic testing Authors: CAO Jing-rong SHEN Ding-Xia Qian Miu LUO Yan-ping Xu Yaping Tian Fang 【Abstract】 Objective To study the People’s Liberation Army General Hospital of multiple drug-resistant clinical isolates of Pseudomonas aeruginosa active efflux system, in order to guide clinical provide a basis for rational choice of antibiotics. Methods efflux pump inhibitor (L-arginine-phenylalanine   β naphthyl amide) and quinolone antibiotics in order to increase micro-broth dilution method with and without efflux pump inhibitor ciprofloxacin and left when the Ofloxacin minimum inhibitory concentration (MIC) values, and calculate the changes in MIC values, according to Canadian and non-Canadian MIC value of efflux pump inhibitors reduce the dilution and over three screening active efflux phenotype positive strains; PCR PCR-positive efflux phenotype of the bacterial resistance gene oprM, mexB and mexR, and three strains for mexR gene sequencing analysis; RT  PCR detection of oprM gene mRNA level, gene expression analysis of drug resistance. The results of phenylalanine-arginine   β naphthlamide can greatly reduce the levofloxacin and ciprofloxacin MIC values of 70 multi-resistant Pseudomonas aeruginosa in the 46 phenotype-positive ( 65.7%), in which PCR detection of oprM, mexB, and mexR gene-positive strains while 42 (91.3%), only the oprM gene positive 2 (4.3%), oprM, mexB, and mexR genes were negative 2 (4.3%). RT  PCR product of measured A value (A260/A280)-positive strains and the PAO1 control 13 to PAO1’s oprM / rpoD value for the control of A-positive strains 4, while meeting the A value (A260/A280) and oprM / rpoD ratio PAO1 strains were higher than 3. Results with the known GenBank sequence of the mexR gene comparison, in which two homologous 100%, a first 434 nucleotide insertion 2bp (AT). Conclusion People’s Liberation Army General Hospital, Most of them are mu