ABSTRACT
BACKGROUND: Colistin has become a last-resort antibiotic for the management of multidrug-resistant gram-negative bacteria. The disk diffusion test is cheap and easy to perform but may be unreliable for colistin susceptibility testing due to poor diffusion of the large colistin molecule. An improved agar diffusion test would increase the reliability of colistin susceptibility testing. This study aimed to modify Muller-Hinton agar (MHA) to improve colistin diffusion in agar.METHODS: MHA was modified by reducing the agar concentration from 100% to 30% and supplementing with protamine. We tested 60 gram-negative clinical isolates of Pseudomonas aeruginosa (N=27) and Acinetobacter calcoaceticus-baumannii complex (N=33). Disk diffusion test results were interpreted based on minimum inhibitory concentrations determined by broth microdilution.RESULTS: The modified MHA yielded the best performance metrics, including 94.7% sensitivity, 100% specificity, and an area under the curve of 0.995 (95% confidence interval, 0.982–1.000), P<0.001, at a cut-off point of 13 mm.CONCLUSIONS: A reduction of the agar concentration from 100% to 30% and the addition of protamine improved colistin diffusion in agar and allowed routine colistin susceptibility testing in a clinical microbiology laboratory, but should be handled with caution.
ABSTRACT
Las infeccionesproducidas por microorganismos multirresistentes son uno de los mayores problemas en los centros asistenciales. Frecuentemente, sólo las polimixinas muestran actividad “in vitro” frente a aislamientos de bacilos gram-negativos resistentes a los carbapenemes. Sin embargo, el National Committee for Clinical Laboratory Standards (NCCLS) no incluye, actualmente, recomendaciones para la realización de las pruebas de sensibilidad para este grupo de antibióticos. Se determinóla actividad de colistín y la correlación entre las pruebas de difusión y dilución de este antibiótico frente a 186 aislamientos contemporáneos en el Hospital de Clínicas “José de San Martín”, siguiendo las recomendaciones generales del NCCLS. Se evaluaron dos puntos de corte: NCCLS 1981 (resistente £ 8 mm y sensible > 11mm) y R. Jones 2001 (resistente £ 11mm y sensible > 14mm). Utilizando el punto de corte del NCCLS 1981 se cometieron los siguientes errores: 0,5% “minor”; 2,2% “major” y 4,4% “very major”, mientras que con el propuesto por R. Jones 2001: 18,9% “minor”; 3,8% “major” y 0,5% “very major”. En conclusión, dado que el punto de corte utilizado por R. Jones 2001 disminuye el error “very major” pero aumenta el “minor” se recomienda la utilización de la concentración inhibitoria mínima (CIM) para confirmar la sensibilidad a colistín cuando sea usada en el tratamiento de infecciones, sin embargo no se detectó resistencia a colistín con halos de inhibición > a 16 mm.
Infections produced by multidrug resistant organisms are one of the greatest problems in health centers. Often, only polymyxines show good activity “in vitro” against the carbapenem resistant gram-negative strains; but the National Committee for Clinical Laboratory Standards (NCCLS) documents do not currently provide interpretative criteria for testing the polymyxines.The antimicrobial activity ofcolistin,and the correlation betweenthe agar dilution test and disk diffusion test were evaluated against 186 gram-negative strains isolated at the Hospital de Clínicas “José de San Martín” of Buenos Aires city. All susceptibility tests were performed according to the NCCLS recommendations. Were evaluated two breakpoints, NCCLS 1981 (£ 8mm and >11mm), and R. Jones 2001 (£ 11 mm and > 14 mm). Discrepancies on interpretative category were found (0.5% minor; 2.2% major and 4.4% very major) with NCCLS 1981, and (18.9% minor; 3.8% majorand 0.5% very major) with R. Jones 2001 criteria. Conclusions. In spite of the fact that the breakpoint used by R. Jones 2001decreases the very major error but increases the minor error, according to our results we recommend the use of MIC methods to assist the therapeutic application of colistin; however resistance to colistin was not detected with zone diameters > 16mm.