Comparison of five methods for detection of carbapenemases in Enterobacterales with proposal of a new algorithm.

Objectives: The aim of this study was to evaluate the performance of five different carbapenemase tests and to develop an algorithm which will permit the detection of most common and rare carbapenemases in routine microbiology laboratories.

Methods: The immunochromatographic tests CARBA-5 (NG), RESIST-4 O.K.N.V. (Coris), the colorimetric β-CARBA (BioRad), a newly developed carbapenem-inactivation method (CIM) supplemented with zinc (zCIM), and the Xpert Carba-R (Cepheid) were challenged with a collection of 189 molecularly characterized Enterobacterales isolates, including 146 carbapenemase producers (CPE): VIM (n = 48), OXA-48-like (n = 40), NDM (n = 29), KPC (n = 13), IMI (n = 9), IMP (n = 9), OXA-58 (n = 2), and GES (n = 2).

Results: The overall sensitivity/specificity values for the five carbapenemase detection tests were 84.2% (CI 77.6-89.2%)/100% (CI 91.8-100%) for RESIST-4, 88.2% (CI 82.1-92.4%)/100% (CI 91.8-100%) for CARBA-5, 88.2% (CI 82.1-92.4%)/100% (CI 91.8-100%) for Xpert Carba-R, 73.7% (CI 66.2-80.0%)/100% (CI 93.4-99.0%) for β-CARBA, and 97.4% (CI 87.9-99.6%)/97.7% (CI 87.9-99.6%) for zCIM. The four common carbapenemases (KPC, OXA-48-like, NDM, and VIM) were detected with ≥97.6% sensitivity by all tests except for β-CARBA (76.6% (CI 68.4-83.2%)). IMI and GES were only detected by zCIM (sensitivity 90.9% (CI 62.3-98.4%)). Based on these results a new algorithm was developed, consisting of an immunochromatographic assay as the first test followed by zCIM, which allows detection of 99.3% of all carbapenemases assessed.

Conclusions: Except for β-CARBA, all methods showed excellent sensitivity/specificity for the detection of the four most frequent carbapenemases. With the new algorithm, rare variants can also be detected. It is rapid, simple, and inexpensive and can be performed in any microbiology laboratory, as no PCR equipment is required.