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Title: Detection of Colistin-Resistant MCR-1-Positive Escherichia coli by Use of Assays Based on Inhibition by EDTA and Zeta Potential
Authors: Esposito, Fernanda
Fernandes, Miriam R.
Lopes, Ralf
Munoz, Maria
Sabino, Caetano P.
Cunha, Marcos P.
Silva, Ketrin C.
Cayo, Rodrigo [UNIFESP]
Martins, Willames M. B. S. [UNIFESP]
Moreno, Andrea M.
Knobl, Terezinha
Gales, Ana C. [UNIFESP]
Lincopan, Nilton
Keywords: polymyxins
colistin resistance
combined disk test
polymyxin NP test
charge modification
Issue Date: 2017
Publisher: Amer Soc Microbiology
Citation: Journal Of Clinical Microbiology. Washington, v. 55, n. 12, p. 3454-3465, 2017.
Abstract: The emergence and rapid dissemination of colistin-resistant Escherichia coli carrying the plasmid-mediated mcr-1 gene have created an urgent need to develop specific screening methods. In this study, we evaluated four assays based on the inhibition of MCR-1 activity by EDTA: (i) a combined-disk test (CDT) comparing the inhibition zones of colistin and colistin (10 mu g) plus EDTA (100 mM); (ii) reduction of colistin MIC (CMR) in the presence of EDTA (80 mu g/ml); (iii) a modified rapid polymyxin Nordmann/Poirel test (MPNP); and (iv) alteration of zeta potential (R-ZP = ZP(+EDTA)/ZP(-EDTA)). We obtained encouraging results for the detection of MCR-1 in E. coli isolates recovered from human, food, and animal samples, using the following assay parameters: >= 3 mm difference in the inhibition zones between colistin disks without and with EDTA; >= 4-fold colistin MIC decrease in the presence of EDTA; R-ZP of >= 2.5; and the absence of metabolic activity and proliferation, indicated by unchanged color of phenol red in the presence of colistin-EDTA, in the MPNP test. In this regard, the CDT, CMR, R-ZP, and MPNP assays exhibited sensitivities of 96.7, 96.7, 95.1, and 96.7% and specificities of 89.6, 83.3, 100, and 100%, respectively, for detecting MCR-1-positive E. coli. Our results demonstrate that inhibition by EDTA and zeta potential assays may provide simple and inexpensive methods for the presumptive detection of MCR-1-producing E. coli isolates in human and veterinary diagnostic laboratories.
ISSN: 0095-1137
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