Author Correction: Rapid susceptibility profiling of carbapenem-resistant Klebsiella pneumoniae

Abstract

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Figure 1

Standardised gating applied to raw data. (A) Collected events were gated to include only those with a SYTO®9 (BL1 - 530/30 nm) fluorescence of 10⁴ arbitrary fluorescence units or higher. Doublets were removed via a FSC-A vs FSC-H plot. Background was removed by plotting specific SYTO®9 fluorescence (BL1 – 530/30) against an unused channel (BL3 – 640 LP). (B) In the antibiotic unexposed sample, 10% nearest-neighbour contouring was applied, and a gate (referred to as Unexposed Cell Morphotype) was set to include all clustered events. This gate was then applied to all samples across the antibiotic dilution series.

Figure 2

Susceptibility to meropenem can be identified by AFC by observing a susceptibility-associated signature. (A) Exposure of the K. pneumoniae susceptible type strain (ATCC 700603) increased forward scatter, SYTO®9 fluorescence, reduced overall event numbers, and formed a new contouring focus at the isolate’s MIC (0.25 mg/L). At 32 × MIC, a total of four contouring foci were observed, with an overall shift towards low forward scatter, low fluorescent debris. The progression of these features, when observed in combination, constitutes the susceptibility-associated signature. Colouring on biaxial plots indicates separate contouring foci. Fluorescence micrographs (acquired at 60x magnification) show reduced overall cell numbers and increase aberrant cell morphotypes as meropenem concentration increases. (B) Exposure of highly resistant clinical K. pneumoniae strain K8 to meropenem shows an absence of susceptibility-associated signature across clinically relevant meropenem concentrations by flow cytometer bi-axial plot, and an absence of aberrant cell morphotypes by fluorescence microscopy.