A multiplex lateral flow immunoassay for the rapid identification of NDM-, KPC-, IMP- and VIM-type and OXA-48-like carbapenemase-producing enterobacteriaceae

Hervé Boutal (1) , Anais Vogel (1) , Sandrine Bernabeu (2) , Karine Devilliers (1) , Elodie Creton (3, 2) , Garence Cotellon (3, 2) , Marc Plaisance (1) , Saoussen Oueslati (2) , Laurent Dortet (3, 2) , Agnès Jousset (3, 2) , Stéphanie Simon (4, 2) , Thierry Naas (3, 2) , Hervé Volland (1)

1 INRA - Institut National de la Recherche Agronomique
2 Université Paris Saclay (COmUE)
3 CNR - Centre National de Référence de la Résistance aux Antibiotiques
4 SPI - Service de Pharmacologie et Immunoanalyse

Objectives: The global spread of carbapenemase-producing Enterobacteriaceae represents a substantial challenge in clinical practice and rapid and reliable detection of these organisms is essential. The aim of this study was to develop and validate a lateral flow immunoassay (Carba5) for the detection of the five main carbapenemases (KPC-, NDM-, VIM-and IMP-type and OXA-48-like). Methods: Carba5 was retrospectively and prospectively evaluated using 296 enterobacterial isolates from agar culture. An isolated colony was suspended in extraction buffer and then loaded on the manufactured Carba5. Results: All 185 isolates expressing a carbapenemase related to one of the Carba5 targets were correctly and unambiguously detected in <15 min. All other isolates gave negative results except those producing OXA-163 and OXA-405, which are considered low-activity carbapenemases. No cross-reaction was observed with non-targeted carbapenemases, ESBLs, AmpCs or oxacillinases (OXA-1, -2, -9 and -10). Overall, this assay reached 100% sensitivity and 95.3%(retrospectively) to 100% (prospectively) specificity. Conclusions: Carba5 is efficient, rapid and easy to implement in the routine workflow of a clinical microbiology laboratory for confirmation of the five main carbapenemases encountered in Enterobacteriaceae.

Mots clés: carba np test; desorption ionization-time; flight mass-spectrometry; k-set assay; immunochromatographic test; multicenter evaluation; cultured bacteria; beta-lactamase; pcr

Domaines: Sciences du Vivant [q-bio]

Hervé Boutal, Anais Vogel, Sandrine Bernabeu, Karine Devilliers, Elodie Creton, et al.. A multiplex lateral flow immunoassay for the rapid identification of NDM-, KPC-, IMP- and VIM-type and OXA-48-like carbapenemase-producing enterobacteriaceae. Journal of Antimicrobial Chemotherapy, 2018, 73 (4), pp.909-915. ⟨10.1093/jac/dkx521⟩. ⟨hal-02629307⟩