Colistin resistance in carbapenem non-susceptible Acinetobacter baumanii in a tertiary care hospital in India: clinical characteristics, antibiotic susceptibility and molecular characterization.

INTRODUCTION: Acinetobacter baumanii (AB) is a bacterium of concern in the hospital setup due to its ability to thrive in unfavorable conditions and the rapid emergence of antibiotic resistance. Carbapenem resistance in this organism is disheartening, further clouded by the emergence of colistin resistance. AIM: The present prospective study aims to note the epidemiology, molecular profile, and clinical outcome of patients with colistin resistance AB infections in a multispecialty tertiary care setup in Odisha, Eastern India. METHODS: All AB strains received from March 2021 to February 2022, identified by Vitek2 (Biomerieux) and confirmed by oxa-51 genes, were included. Carbapenem and colistin resistance were identified as per CLSI guidelines. Known mutations for blaOXA-23-like, blaIMP, blaVIM, blaKP, lpxA, lpxC, pmrA, pmrB, and plasmid mediated mcr (mcr1-5) were screened by conventional PCR techniques. The clinical outcome was noted retrospectively from case sheets. Data was entered in MS Excel and tabulated using SPSS software. RESULTS: In the study period, 350 AB were obtained, of which 317(90.5%) were carbapenem resistant (CRAB). Among the CRAB isolates, 19 (5.9%) were colistin resistant (ABCoR). The most valuable antibiotics in the study were tigecycline (65.4% in ABCoI; 31.6% in ABCoR) and minocycline (44.3% in CI; 36.8% in CR). There was a significant difference in mortality among ABCoI and ABCoR infections. bla OXA was the predominant carbapenem resistance genotype, while pmrA was the predominant colistin resistant genotype. There were no plasmid mediated mcr genes detected in the present study.

Evaluation of different methods for in vitro susceptibility testing of colistin in carbapenem resistant Gram-negative bacilli.

Introduction: The increasing antibiotic resistance like the advent of carbapenem resistant Enterobactarales (CRE), Carbapenem Resistant Acinetobacter baumanii (CRAB), and Carbapenem Resistant Pseudomonas aeruginosa (CRPA) has led to to the use of toxic and older drugs like colistin for these organisms. But worldwide there is an increase in resistance even to colistin mediated both by chromosomes and plasmids. This necessitates accurate detection of resistance. This is impeded by the unavailability of a user-friendly phenotypic methods for use in routine clinical microbiology practice. The present study attempts to evaluate two different methods - colistin broth disc elution and MIC detection by Vitek two in comparison to CLSI approved broth microdilution (BMD) for colistin for Enterobactarales, Pseudomonas aeruginosa , and Acinetobacter baumanii clinical isolates. Methods: Colistin susceptibility of 6013 carbapenem resistant isolates was determined by BMD, Colistin Broth Disc Elution (CBDE), and Vitek two methods and was interpreted as per CLSI guidelines. The MIC results of CBDE, Vitek two were compared with that of BMD and essential agreement (EA), categorical agreement (CA), sensitivity, specificity, very major error (VME), major error (ME) and Cohen's kappa (CK) was calculated. The presence of any plasmid-mediated colistin resistance (mcr-1, 2, 3, 4 and 5) was evaluated in all colistin-resistant isolates by conventional polymerase chain reaction. Results: Colistin resistance was found in 778 (12.9 %) strains among the carbapenem resistant isolates. Klebsiella pneumoniae had the highest (18.9 %) colistin resistance by the BMD method. MIC of Vitek two had sensitivity ranging from 78.2-84.8% and specificity of >92 %. There were 171 VMEs and 323 MEs by Vitek two method, much more than CLSI acceptable range. The highest percentage of errors was committed for Acinetobacter baumanii (27.8 % of VME and 7.9 % ME). On the other hand, the CBDE method performed well with EA, CA, VME and ME within acceptable range for all the organisms. The sensitivity of the CBDE method compared to gold standard BMD varied from 97.5-98.8 % for different strains with a specificity of more than 97.6 %. None of the isolated colistin resistant organisms harboured mcr plasmids. Conclusion: As BMD has many technical complexities, CBDE is the best viable alternative available for countries like India. A sensitive MIC reported by Vitek two needs to be carefully considered due high propensity for VMEs particularly for Klebsiella spp.