Artigo

Novel Cassette Assay To Quantify the Outer Membrane Permeability of Five B-Lactams Simultaneously in Carbapenem-Resistant Klebsiella pneumoniae and Enterobacter cloacae

Bartolome Moya, Yinzhi Lang, Alexandre Prehn Zavascki, Tae Hwan Kim, Xun Tao, Yuanyuan Jiao, Kari B. Basso, Jieqiang Zhou, Dhruvitkumar S. Sutaria, Afonso L. Barth, Stephanie M. Reeve

American Society for Microbiology, v. 11, n. 1, p. 1-15, 2020.

Motivo: Professor IEP

Área da saúde: Medicina, Medicina - Acupuntura, Medicina - Anestesiologia, Medicina - Angiologia, Medicina - Cardiologia, Medicina - Cirurgia Aparelho Digestivo, Medicina - Cirurgia Cardiovascular, Medicina - Cirurgia Geral, Medicina - Cirurgia Pediátrica, Medicina - Cirurgia Plástica, Medicina - Cirurgia Torácica, Medicina - Cirurgia Vascular, Medicina - Cirurgia da Mão, Medicina - Cirurgia de Cabeca e Pescoco, Medicina - Clinica Medica, Medicina - Coloproctologia, Medicina - Dermatologia, Medicina - Endocrinologia e Metabologia, Medicina - Endoscopia, Medicina - Epidemiologia, Medicina - Fisioterapia, Medicina - Física de Radioterapia, Medicina - Gastroenterologia, Medicina - Genética Medica, Medicina - Geriatria, Medicina - Ginecologia e Obstetrícia, Medicina - Hematologia e Hemoterapia, Medicina - Infectologia, Medicina - Mastologia, Medicina - Medicina Do Trabalho, Medicina - Medicina Física e Reabilitação, Medicina - Medicina Intensiva, Medicina - Medicina Nuclear, Medicina - Medicina de Família e Comunidade, Medicina - Nefrologia, Medicina - Neurocirurgia, Medicina - Neurologia, Medicina - Oftalmologia, Medicina - Oncologia, Medicina - Ortopedia e Traumatologia, Medicina - Otorrinolaringologia, Medicina - Patologia, Medicina - Patologia Clinica/Medicina Laboratorial, Medicina - Pediatria, Medicina - Pneumologia, Medicina - Psicologia, Medicina - Psiquiatria, Medicina - Radiologia e Diagnóstico por Imagem, Medicina - Radioterapia, Medicina - Reumatologia, Medicina - Terapia Intensiva, Medicina - Urologia

Resumo: Poor penetration through the outer membrane (OM) of Gram-negative bacteria is a major barrier of antibiotic development. While B-lactam antibiotics are commonly used against Klebsiella pneumoniae and Enterobacter cloacae, there are limited data on OM permeability especially in K. pneumoniae. Here, we developed a novel cassette assay, which can simultaneously quantify the OM permeability to five B-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Both clinical isolates harbored a blaKPC-2 and several other B-lactamases. The OM permeability of each antibiotic was studied separately (“discrete assay”) and simultaneously (“cassette assay”) by determining the degradation of extracellular B-lactam concentrations via multiplex liquid chromatography-tandem mass spectrometry analyses. Our K. pneumoniae isolate was polymyxin resistant, whereas the E. cloacae was polymyxin susceptible. Imipenem penetrated the OM at least 7-fold faster than meropenem for both isolates. Imipenem penetrated E. cloacae at least 258-fold faster and K. pneumoniae 150-fold faster compared to aztreonam, cefepime, and ceftazidime. For our B-lactams, OM permeability was substantially higher in the E. cloacae compared to the K. pneumoniae isolate (except for aztreonam). This correlated with a higher OmpC porin production in E. cloacae, as determined by proteomics. The cassette and discrete assays showed comparable results, suggesting limited or no competition during influx through OM porins. This cassette assay allowed us, for the first time, to efficiently quantify the OM permeability of multiple B-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Characterizing the OM permeability presents a critical contribution to combating the antimicrobial resistance crisis and enables us to rationally optimize the use of B-lactam

Link: https://journals.asm.org/doi/10.1128/mBio.03189-19

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