2022Klimik Dergisi/Klimik JournalOpen access

Evaluation of Colistin Susceptibility Results in Multidrug-Resistant Gram-Negative Bacilli by Colistin Broth Disk Elution Method

Ayşe Özkaçmaz, Fikriye Milletli Sezgin

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Abstract

Objective: Determining colistin susceptibility according to the reference method is a significant problem for clinical microbiology laboratories. There is a need for a fast, easy-to-apply, and accurate method for laboratories with limited opportunities. In our study, we aimed to evaluate the applicability of the colistin broth disk elution (CBDE) test. Methods: A total of 193 Gram-negative bacteria, which were isolated from various clinical specimens in our laboratory, were included in the study. Colistin susceptibility was determined by broth microdilution as the reference method. The CBDE test was performed as previously described. For each bacterial isolate, 0, ½, 1, 2, and 4 colistin discs (10 µg; BD) were added to 5 tubes containing cation-adjusted Mueller-Hinton broth (MHB). The final colistin concentrations in the tubes were 0 (growth control), 0.5, 1, 2, and 4 µg/ml, respectively. The minimum inhibitory concentration values were determined by visual evaluation 16-20 hours after the bacterial inoculation. Results: In the Enterobacterales group, the categorical agreement was 94%, the major error was 5%, and the very major error was 0%. In the non-fermentative bacteria group, the categorical agreement was found to be 94%, the major error 1%, and the very major error 33%, respectively. Conclusion: The CBDE test is a good alternative to broth microdilution in routine microbiology laboratories for the Enterobacterales group since it provides easy-to-apply, cost-effective testing for colistin susceptibility and has a high categorical agreement compared to the reference microdilution method. Keywords: colistin, disc elution, antibiotic resistance, broth microdilution, Enterobacterales

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Objective: Determining colistin susceptibility according to the reference method is a significant problem for clinical microbiology laboratories. There is a need for a fast, easy-to-apply, and accurate method for laboratories with limited opportunities. In our study, we aimed to evaluate the applicability of the colistin broth disk elution (CBDE) test. Methods: A total of 193 Gram-negative bacteria, which were isolated from various clinical specimens in our laboratory, were included in the study. Colistin susceptibility was determined by broth microdilution as the reference method. The CBDE test was performed as previously described. For each bacterial isolate, 0, ½, 1, 2, and 4 colistin discs (10 µg; BD) were added to 5 tubes containing cation-adjusted Mueller-Hinton broth (MHB). The final colistin concentrations in the tubes were 0 (growth control), 0.5, 1, 2, and 4 µg/ml, respectively. The minimum inhibitory concentration values were determined by visual evaluation 16-20 hours after the bacterial inoculation. Results: In the Enterobacterales group, the categorical agreement was 94%, the major error was 5%, and the very major error was 0%. In the non-fermentative bacteria group, the categorical agreement was found to be 94%, the major error 1%, and the very major error 33%, respectively. Conclusion: The CBDE test is a good alternative to broth microdilution in routine microbiology laboratories for the Enterobacterales group since it provides easy-to-apply, cost-effective testing for colistin susceptibility and has a high categorical agreement compared to the reference microdilution method. Keywords: colistin, disc elution, antibiotic resistance, broth microdilution, Enterobacterales

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Available abstract

Objective: Determining colistin susceptibility according to the reference method is a significant problem for clinical microbiology laboratories. There is a need for a fast, easy-to-apply, and accurate method for laboratories with limited opportunities. In our study, we aimed to evaluate the applicability of the colistin broth disk elution (CBDE) test. Methods: A total of 193 Gram-negative bacteria, which were isolated from various clinical specimens in our laboratory, were included in the study. Colistin susceptibility was determined by broth microdilution as the reference method. The CBDE test was performed as previously described. For each bacterial isolate, 0, ½, 1, 2, and 4 colistin discs (10 µg; BD) were added to 5 tubes containing cation-adjusted Mueller-Hinton broth (MHB). The final colistin concentrations in the tubes were 0 (growth control), 0.5, 1, 2, and 4 µg/ml, respectively. The minimum inhibitory concentration values were determined by visual evaluation 16-20 hours after the bacterial inoculation. Results: In the Enterobacterales group, the categorical agreement was 94%, the major error was 5%, and the very major error was 0%. In the non-fermentative bacteria group, the categorical agreement was found to be 94%, the major error 1%, and the very major error 33%, respectively. Conclusion: The CBDE test is a good alternative to broth microdilution in routine microbiology laboratories for the Enterobacterales group since it provides easy-to-apply, cost-effective testing for colistin susceptibility and has a high categorical agreement compared to the reference microdilution method. Keywords: colistin, disc elution, antibiotic resistance, broth microdilution, Enterobacterales

Key concepts: Colistin, Broth microdilution, Minimum inhibitory concentration, Microbiology, Bacilli, Biology, Bacteria, Antibiotics

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