Abstract
ABSTRACTThe global spread of carbapenemase-producing Enterobacteriaceae (CPE) that are often resistant to most, if not all, classes of antibiotics is a major public health concern. The NDM-1 carbapenemase is among the most worrisome carbapenemases given its rapid worldwide spread. We have developed and evaluated a lateral flow immunoassay (LFIA) (called the NDM LFIA) for the rapid and reliable detection of NDM-like carbapenemase-producing Enterobacteriaceae from culture colonies. We evaluated the NDM LFIA using 175 reference enterobacterial isolates with characterized β-lactamase gene content and 74 nonduplicate consecutive carbapenem-resistant clinical isolates referred for expertise to the French National Reference Center (NRC) for Antibiotic Resistance during a 1-week period (in June 2016). The reference collection included 55 non-carbapenemase producers and 120 carbapenemase producers, including 27 NDM producers. All 27 NDM-like carbapenemase producers of the reference collection were correctly detected in less than 15 min by the NDM LFIA, including 22 strains producing NDM-1, 2 producing NDM-4, 1 producing NDM-5, 1 producing NDM-7, and 1 producing NDM-9. All non-NDM-1 producers gave a negative result with the NDM LFIA. No cross-reaction was observed with carbapenemases (VIM, IMP, NDM, KPC, and OXA-48-like), extended-spectrum β-lactamases (ESBLs) (TEM, SHV, and CTX-M), AmpCs (CMY-2, DHA-2, and ACC-1), and oxacillinases (OXA-1, -2, -9, and -10). Similarly, among the 74 referred nonduplicate consecutive clinical isolates, all 7 NDM-like producers were identified. Overall, the sensitivity and specificity of the assay were 100% for NDM-like carbapenemase detection with strains cultured on agar. The NDM LFIA was efficient, rapid, and easy to implement in the routine workflow of a clinical microbiology laboratory for the confirmation of NDM-like carbapenemase-producing Enterobacteriaceae.
Highlights
The global spread of carbapenemase-producing Enterobacteriaceae (CPE) that are often resistant to most, if not all, classes of antibiotics is a major public health concern
ESBL-producing Enterobacteriaceae are resistant to all -lactams up to third-generation cephalosporins [2], and this leads to the use of last-resort antibiotics such as carbapenems
Isolated in India, it is endemic in the entire Indian subcontinent and represents a major threat considering the high rate of acquisition (72.4%) of multiresistant Enterobacteriaceae (MRE), including NDM-1-expressing strains, while traveling in this area [8]
Summary
The global spread of carbapenemase-producing Enterobacteriaceae (CPE) that are often resistant to most, if not all, classes of antibiotics is a major public health concern. A lateral flow immunoassay (LFIA) for the detection of OXA-48-type carbapenemases has been evaluated [18] This test yields results from cultured strains within 15 min, with 100% specificity and sensitivity. To respond to the needs optimally, antimicrobial drug resistance detection methods must be cheap (reduced costs of consumables and equipment) and easy to use (reduced technical complexity) for the end user This led us to develop an LFIA for the detection of NDM-like carbapenemases that presents several advantages: robust technology, transferable in a commercialized version, stable for more than 24 months without refrigeration, user-friendly (no requirement for trained staff), high performance (sensitive and specific), low cost (around 6€ per test [22, 23], compared to more than 20€ for molecular tests), and detection results in less than 30 min without the need for highly technical equipment for the readout. We have validated this assay on 175 agar-cultured enterobacterial isolates (27 expressing NDM-like enzymes), with test results in less than 15 min, 100% sensitivity, and 100% specificity
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