Rapid detection of carbapenemase-producing Klebsiella pneumoniae: A comparative study of CIM and MHT

Year 2025, Volume: 38 Issue: 1, 49 - 51, 29.01.2025

Fatih Mehmet Akıllı , Ege Kuyumcu Burak Kurt Muhammet Talha Korucu , Hüseyin Nafiz Uysal Kaan Sayan Arzu Akşit İlki

https://doi.org/10.5472/marumj.1627923

Abstract

Objective: This study aimed to compare the performance of the carbapenem inactivation method (CIM) and the modified hodge test
(MHT) to screen carbapenemase activity in Klebsiella pneumoniae isolates with genotypically confirmed results.
Materials and Methods: A total of 114 carbapenem-resistant K. pneumoniae clinical isolates were collected from hospitalized patients.
Two methods, the MHT and CIM, were used to investigate carbapenemase production. The CIM test was evaluated at the 6th hour for
the preliminary decision and the 24th hour for the final decision. MHT was evaluated at the 24th hour. A polymerase chain reaction
(PCR) was performed to detect carbapenemase-encoding genes (blaKPC, blaNDM, blaIMP, blaVIM, and blaOXA-48).
Results: Of these isolates in which a carbapenemase enzyme was detected by PCR, blaOXA-48 was found in 87.7%, blaKPC in 6.1%, blaNDM
in 6.1%, blaIMP in 0.8%, and blaVIM in 0.8%. The most common carbapenemase gene detected was OXA-48. Of the 114 isolates with
a genotypically detected carbapenemase enzyme, 98 and 109 were positive by CIM at the 6th and 24th hour, respectively, and 88 by
MHT. The isolates producing both blaOXA-48 and blaNDM and blaIMP and blaVIM were detected as positive by both phenotypic tests. The
sensitivity of CIM at the 6th and 24th hour and MHT was found to be 85.9%, 95.6%, and 77.1%, respectively.
Conclusion: These findings indicate that CIM can be an effective method for accurately and rapidly detecting carbapenemase activity
in K. pneumoniae infections, particularly in clinical microbiological laboratories with limited resources. To verify the negative tests,
molecular methods are recommended to predict OXA-48 activity particularly..

Keywords

Carbapenem-resistant, Carbapenemase, OXA-48, Beta lactamase KPC, PCR

References

• Santajit S, Indrawattana N. Mechanisms of antimicrobial resistance in eskape pathogens. Biomed Res Int 2016;2016:2475067. doi: 10.1155/2016/2475067.
• Das BJ, Singha KM, Wangkheimayum J, Chanda DD, Bhattacharjee A. Emergence of carbapenem-resistant enterobacterales co-harboring blaOXA-78 and blaOXA-58 from India. Ann Clin Microbiol Antimicrob 2023;22:79. doi: 10.1186/s12941.023.00635-6.
• Partridge SR, Kwong SM, Firth N, Jensen SO. Mobile genetic elements associated with antimicrobial resistance. Clin Microbiol Rev 2018;31:e00088-17. doi: 10.1128/CMR.00088- 17.
• Saliba R, Aho-Glélé LS, Karam-Sarkis D, Zahar JR. Evaluation of polymerase chain reaction assays for direct screening of carbapenemase-producing Enterobacteriaceae from rectal swabs: a diagnostic meta-analysis. J Hosp Infect 2020;104:381- 9. doi: 10.1016/j.jhin.2019.11.017
• Nieto-Saucedo JR, López-Jacome LE, Franco-Cendejas R, et al. Carbapenem-resistant gram-negative bacilli characterization in a tertiary care center from El Bajio, Mexico. Antibiotics (Basel) 2023;12:1295. doi: 10.3390/antibiotics12081295.
• Tamma PD, Simner PJ. Phenotypic detection of carbapenemase-producing organisms from clinical isolates. J Clin Microbiol 2018 ;56:e01140-18. doi: 10.1128/JCM.01140- 18
• Haji SH, Aka STH, Ali FA. Prevalence and characterisation of carbapenemase encoding genes in multidrug-resistant Gramnegative bacilli. PLoS One 2021;16:e0259005. doi: 10.1371/ journal.pone.0259005.
• Kakoullis L, Papachristodoulou E, Chra P, Panos G. Mechanisms of antibiotic resistance in important grampositive and gram-negative pathogens and novel antibiotic solutions. Antibiotics (Basel) 2021 ;10:415. doi: 10.3390/ antibiotics10040415.
• Rabaan AA, Eljaaly K, Alhumaid S, et al. An overview on phenotypic and genotypic characterisation of carbapenemresistant enterobacterales. Medicina (Kaunas) 2022;58:1675. doi: 10.3390/medicina58111675.
• van der Zwaluw K, de Haan A, Pluister GN, Bootsma HJ, de Neeling AJ, Schouls LM. The carbapenem inactivation method (CIM), a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods. PLoS One 2015;10:e0123690. doi: 10.1371/journal. pone.0123690.
• Schaffarczyk L, Noster J, Stelzer Y, Sattler J, Gatermann S, Hamprecht A. Detection of rare carbapenemases in Enterobacterales-comparison of two colorimetric and three CIM-based carbapenemase assays. Microbiol Spectr 2024;12:e0301523. doi: 10.1128/spectrum.03015-23.
• Lee YL, Chen HM, Hii IM, Hsueh PR. Carbapenemaseproducing Enterobacterales infections: recent advances in diagnosis and treatment. Int J Antimicrob Agents 2022;59:106528. doi: 10.1016/j.ijantimicag.2022.106528.
• Rizvi M, Sami H, Azam M, et al. Reliability of carbapenem inactivation method (CIM) and modified carbapenem inactivation method (mCIM) for detection of OXA-48-like and NDM-1. Indian J Med Microbiol. 2021;39:451-6. doi: 10.1016/j.ijmmb.2021.07.004.
• Codjoe FS, Donkor ES. Carbapenem resistance: A review. Med Sci (Basel) 2017;6:1. doi: 10.3390/medsci6010001.
• Yang X, Dong N, Chan EW, Zhang R, Chen S. Carbapenem Resistance-encoding and virulence-encoding conjugative plasmids in Klebsiella pneumoniae. Trends Microbiol 2021;29:65-83. doi: 10.1016/j.tim.2020.04.012.
• Cui X, Zhang H, Du H. Carbapenemases in enterobacteriaceae: detection and antimicrobial Therapy Front Microbiol 2019;10:1823. doi: 10.3389/fmicb.2019.01823.
• Li W, Guo H, Gao Y, et al. Comparative genomic analysis of plasmids harboring blaOXA-48-like genes in Klebsiella pneumoniae. Front Cell Infect Microbiol 2022;12:1082813. doi: 10.3389/fcimb.2022.108.2813.
• Lee YL, Wang WY, Ko WC, Hsueh PR. Global epidemiology and antimicrobial resistance of Enterobacterales harbouring genes encoding OXA-48-like carbapenemases: insights from the results of the Antimicrobial Testing Leadership and Surveillance (ATLAS) programme 2018-2021. Antimicrob Chemother 2024;79:1581-9. doi: 10.1093/jac/dkae140.
• Candan ED, Aksöz N. Klebsiella pneumoniae: characteristics of carbapenem resistance and virulence factors. Acta Biochim Pol 2015;62:867-74. doi: 10.18388/abp.2015_1148.
• Xu X, Qu F. [Progress in the diagnosis of carbapenemresistant Enterobacteriaceae]. Sheng Wu Gong Cheng Xue Bao. 2018;34:1338-45. doi: 10.13345/j.cjb.170522.
• Akhi MT, Khalili Y, Ghotaslou R, et al. Carbapenem inactivation: a very affordable and highly specific method for phenotypic detection of carbapenemaseproducing Pseudomonas aeruginosa isolates compared with other methods. J Chemother 2017;29:144-9. doi: 10.1080/1120009X.2016.119.9506.
• Fan S, Dai Y, Hou L, Xu Y. Application value of triton X-100 to Modified Hodge Test and Carbapenem Inactivation Method in the detection of acinetobacter baumannii carbapenemase. Infect Drug Resist 2020;13:4283-8. doi: 10.2147/IDR.S281049.
• Sun K, Xu X, Yan J, Zhang L. Evaluation of six phenotypic methods for the detection of carbapenemases in Gram-negative bacteria with characterized resistance mechanisms. Ann Lab Med 2017;37:305-12. doi: 10.3343/alm.2017.37.4.305.
• Saliba R, Aho-Glélé LS, Karam-Sarkis D, Zahar JR. Evaluation of polymerase chain reaction assays for direct screening of carbapenemase-producing Enterobacteriaceae from rectal swabs: a diagnostic meta-analysis. J Hosp Infect 2020;104:381- 9. doi: 10.1016/j.jhin.2019.11.017
• Kanj SS, Bassetti M, Kiratisin P, et al. Clinical data from studies involving novel antibiotics to treat multidrug-resistant Gram-negative bacterial infections. Int J Antimicrob Agents 2022;60:106633. doi: 10.1016/j.ijantimicag.2022.106633.