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Study on the relationship between structural diversity and drug resistance function of macrolide resistance-related genes in Pseudomonas aeruginosa

Study on the relationship between structural diversity and d...

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2019年浙江省医学会耳鼻咽喉头颈外科学学术大会

作者： Qing Chen Liyan Ni The Second Affiliated Hospital and Yuying Children’s Hospital Wenzhou Medical University

Objective In this work,to analyze the resistance mechanism of Pseudomonas aeruginosa to macrolide *** and methods:Pooled genomic DNAs of 262 clinical *** strains were sequenced,and PCR was applied to screen for macrolide resistance gene-positive *** cloning was performed to clone the resistance genes,and minimum inhibitory concentrations(MICs) were tested by the agar dilution ***:only two macrolide resistance genes,mphe and msrE,were identified in the pooled genomic DNA of the 262 *** *** screening results demonstrated that 4 strains(1.53%,4/262) carried both msrE and mphe,and an additional 3 strains(1.15%,3/262) carried only *** assay results showed that both the cloned msrE and mphe genes(pUCP18-mphe/DH5α and pUCP18-msrE/DH5α) were functional and mediated higher resistance levels to three second-generation macrolides(MIC levels increased≥ 4-fold to roxithromycin,clarithromycin and azithromycin compared with those of the recipient,DH5α) than to two first-generation macrolides(MIC levels increased≤2-fold to erythromycin and kitasamycin compared with those of the recipient,DH5α).Both the mphe and msrE genes conferred the highest resistance to roxithromycin(MICs:1024 μg/mL),but they showed the least resistance to azithromycin(MICs:8 μg/mL and 32 μg/mL,respectively).Analysis of the polymorphism of the proteins encoded by the msrE and mphe genes revealed that among the 6 mphe and 9 msrE variants(including two mphe and two msrE sequences from this work,while the remainder were obtained from a database),the two genes are very *** genes had one to three amino acid differences compared with the reference sequences for both genes(MG585957.1 for msrE and CP029638.1 for mphe).Conclusion:It can be concluded that the known macrolide resistance genes are seldom identified in clinical *** strains isolated in Wenzhou,China,despite several mphe and msrE genes being present in some ***

关键词： Pseudomonas aeruginosa macrolide resistance gene mphe msrE

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