Modification of carbapenemase inhibition test and comparison of its performance with NG-Test CARBA 5 for detection of carbapenemase-producing Enterobacterales

J Appl Microbiol. 2024 Aug 5;135(8):lxae197. doi: 10.1093/jambio/lxae197.

Abstract

Aims: Adequately and accurately identifying carbapenemase-producing Enterobacterales (CPE) is vital for selecting appropriate antimicrobial therapy and implementing effective infection control measures. This study aims to optimize the phenotypic detection method of carbapenemase for routine diagnostics in clinical microbiology laboratories.

Methods and results: Carbapenemase genes in 2665 non-duplicate CRE clinical strains collected from various regions of China were confirmed through whole-genome sequencing (WGS). The carbapenemase inhibition test (CIT) was conducted and interpreted using different methods and breakpoints, then compared with the NG-Test CARBA 5 for carbapenemase detection. The diagnostic performance of the CIT method was optimal when the carbapenemase types were determined by comparing the inhibition zone diameters of the imipenem disc with 3-aminophenylboronic acid (APB) plus ethylenediaminetetraacetic acid (EDTA) to those of the imipenem disc with either APB or EDTA alone, with a breakpoint of 4 mm. The overall sensitivities of the current CIT, the modified CIT, and NG-Test CARBA 5 were 91.4%, 94.9%, and 99.9%, respectively. For detecting isolates co-producing Klebsiella pneumoniae carbapenemase (KPC) and metallo-β-lactamases (MBLs), the modified CIT method had higher sensitivity than the current method (70.0% vs. 53.3%), though this difference was not statistically significant (P = 0.063). The NG-Test CARBA 5 showed excellent performance for multi-carbapenemases diagnosis, with sensitivity and specificity of 97.1% and 100%, respectively.

Conclusions: Optimizing and standardizing the CIT method for clinical use is necessary. It has certain advantages in diagnosing multi-carbapenemase and rare carbapenemase production. However, for identifying common carbapenemase types, the NG-Test CARBA 5 demonstrated superior performance.

Keywords: KPC; NG-Test CARBA 5; carbapenemase inhibition test; carbapenemase-producing Enterobacterales; lateral flow immunochromatographic assay; metallo-β-lactamases; multi-carbapenemase; whole-genome sequencing.

Publication types

  • Comparative Study

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins* / metabolism
  • Carbapenem-Resistant Enterobacteriaceae / drug effects
  • Carbapenem-Resistant Enterobacteriaceae / enzymology
  • Carbapenem-Resistant Enterobacteriaceae / isolation & purification
  • China
  • Enterobacteriaceae / drug effects
  • Enterobacteriaceae / enzymology
  • Enterobacteriaceae / isolation & purification
  • Enterobacteriaceae Infections / microbiology
  • Humans
  • Imipenem / pharmacology
  • Microbial Sensitivity Tests
  • Sensitivity and Specificity
  • Whole Genome Sequencing
  • beta-Lactamases* / analysis
  • beta-Lactamases* / metabolism

Substances

  • beta-Lactamases
  • carbapenemase
  • Bacterial Proteins
  • Imipenem
  • Anti-Bacterial Agents