Evaluation of the Amplidiag CarbaR+VRE Kit for Accurate Detection of Carbapenemase-Producing Bacteria

Abstract

As carbapenemase-producing Gram-negative bacilli (CP-GNB) (, , and spp.) are becoming a major public health issue, there is an urgent need for accurate and fast diagnostic tests. The Amplidiag CarbaR+VRE assay is a multiplex nucleic acid-based diagnostic test intended for the detection of CP-GNB and vancomycin-resistant enterococci (VRE) from cultured colonies. We have evaluated its ability to detect carbapenemase genes in 100 well-characterized GNB and in 200 consecutive enterobacterial isolates with reduced susceptibility to carbapenems that were referred to the French National Reference Center for carbapenem resistance. The assay has been validated on purified DNA but also directly on colonies. The Amplidiag CarbaR+VRE assay could detect all KPC, NDM, VIM, IMP, and OXA-48-like variants tested and all acquired carbapenem-hydrolyzing oxacillinases from (OXA-23, OXA-24/-40, and OXA-58) as well as the overexpressed chromosomally encoded OXA-51-like β-lactamase associated with an upstream inserted IS However, as claimed by the manufacturer, other carbapenemases such as GES-like carbapenemases (GES-2, GES-5, and GES-14), GIM-1, AIM-1, SPM-1, DIM-1, OXA-198 in , or OXA-143-like in were not detected. Amplidiag CarbaR+VRE's performance values were high (100% sensitivity and 99% specificity) as it could detect the five major carbapenemases-NDM, VIM, IMP, KPC, and OXA-48-as well as OXA-type carbapenemases from spp. that are currently emerging also among and other enterobacterial isolates. It can provide a result directly from colonies growing on Mueller-Hinton (MH) agar or on selective screening medium in less than 2 h. Further evaluations are now necessary to determine the performance values directly on rectal swabs.

Detection of Multidrug-Resistant Enterobacterales—From ESBLs to Carbapenemases

“…The predecessor Check-MDR CT103 performed well in the study by Cuzon et al, with sensitivity and specificity of 95–100% and 100%, respectively [ 84 ]. Other tests for CPE detection from rectal swabs include the Xpert Carba-R assay for the GeneXpert system (Cepheid, Sunnyvale, California [ 139 , 140 , 141 ]), the CRE ELITe MGB Kit (ELITechGroup, Puteaux, France [ 85 ]), GenePOC/Revogene Carba C assay (Meridian Bioscience, Cincinnati, Ohio [ 46 ]), and assays developed by MOBIDIAG for the Amplidiag ® Easy system, namely, Amplidiag CarbaR + VRE and Amplidiag CarbaR-MCR assays (Mobidiag Ltd., Espoo, Finland [ 86 , 87 ]).…”

Evaluation of the Novodiag CarbaR+, a Novel Integrated Sample to Result Platform for the Multiplex Qualitative Detection of Carbapenem and Colistin Resistance Markers

“…Several other assays are commercially available for that purposes. Some require DNA extraction or at least simple sample heating of rectal swabs specimen before molecular analysis (PCR or isothermal Loop-mediated amplification [LAMP]) such as PCR-based Amplidiag Ò CARBA+MCR assay (Mobidiag, Paris, France), 41 Amplidiag CAR-BA+VRE assay (Mobidiag), 42 the Check-Direct CPE assay (Check-Points, Wageningen, The Netherlands), 43 or LAMP easyplex Ò SuperBug panel (Amplex, Gars-Bahnhof, Germany). 44,45 Others are fully automated, such as CRE ELITe MGB Ò kits on an InGenius platform (Elitech, Les Ulis, France), 46 BD MAXÔ CPO assay (Becton-Dickinson, Le Pont-de-Claix, France), 47 and Xpert Carba-R assay (Cepheid, Sunnyvale, CA).…”

Evaluation of the Revogene Carba C Assay for Detection and Differentiation of Carbapenemase-Producing Gram-Negative Bacteria

“…Several assays are commercially available. Some require DNA extraction from rectal swab specimens prior to amplification, such as the PCR-based Amplidiag CARBAϩMCR assay (Mobidiag, Paris, France) (24), the Amplidiag CARBAϩVRE assay (Mobidiag, Paris, France) (25), or the Check-Direct CPE assay (Check-Points, Wageningen, The Netherlands) (26), while others do not require DNA extraction, such as the Eazyyplex SuperBug CRE (Amplex Biosystems GmbH, Gießen, Germany) (27). Others are fully automated, such as CRE ELITe MGB kits on the InGenius platform (Elitech, Les Ulis, France) (28), the BD MAX Checkpoint CPO assay (29), and the GeneXpert Carba-R assay (Cepheid, Sunnyvale, CA, USA) (30,31).…”