PmrA-PmrB二元调控系统介导大肠杆菌对黏杆菌素耐药的机制研究

doi:10.11843/j.issn.0366-6964.2016.04.022

Abstract

Abstract:

It has been recently reported that PmrA-PmrB are associated with polymyxin B resistance.To determine whether PmrA-PmrB is contributing to colistin resistance in E.coli，we investigated amino acid alterations and transcription level of pmrA-pmrB in colistin resistance strains.All clinical resistance isolated strains were screened for colistin resistance by using MIC determinations and step-wise method was used to induce colistin resistant strains. pmrA-pmrB genes were amplified by PCR from resistant strain and their sequences were characterized by Mega software.Simultaneously，expression levels of PmrA-PmrB were detected by quantitive Real-time PCR.MIC (minimum inhibitory concentration) results showed that 88.5% of 52 clinic resistance strains tested in this study were susceptibility to colistin and 11.5% of the strains were resistant.Five mutations were identified in pmrB in induced colistin-resistant mutants 9R (G55A)，36R (T500C)，53R (T263A)，91R (insertion of 30 bp at position of 229) and 107R (insertion of 189 bp at position of 478).However，six clinical resistance isolates (MIC=4-8 μg•mL^-1) did not show any mutations in pmrA-pmrB.Further study showed that pmrA-B expression levels were significantly activated in three lab-derived colistin resistant strains with point mutations of pmrB.The expression level of pmrA-pmrB genes tended to increase but not significant changed in resistant strains with insertion mutations compared with its correspond parent strains.The transcription level of pmrA-pmrB in 6 clinical resistance isolates remained unchanged compared with susceptible strains.In addition，when MICs were over than 16 μg•mL^-1，mutations in pmrA-pmrB would occur.Mutations in pmrB or high expression levels in pmrA-pmrB contribute to high-level colistin resistance in E.coli strains.

CLC Number:

S859.7

GE Lin，GUO Da-wei，HE Fang，HUANG Jin-hu，WANG Li-ping. Resistance Mechanism of Escherichia coli to Colistin Mediated by PmrA-PmrB[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 812-819.

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Resistance Mechanism of Escherichia coli to Colistin Mediated by PmrA-PmrB

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