Performance of a Novel Fluorogenic Assay for Detection of Carbapenemase-Producing Enterobacteriaceae from Bacterial Colonies and Directly from Positive Blood Cultures
- Authors
- Kim, Hoon Seok; Kim, Jung Ok; Lee, Ji Eun; Park, Kang Gyun; Lee, Hae Kyung; Kim, Soo-Young; Min, Sun-Joon; Kim, Juhyeon; Park, Yeon-Joon
- Issue Date
- Jan-2020
- Publisher
- American Society for Microbiology
- Keywords
- fluorogenic assay; carbapenemase-producing Enterobacteriaceae; positive blood culture; direct; fluorogenic probe
- Citation
- Journal of Clinical Microbiology, v.58, no.1, pp.1 - 11
- Indexed
- SCIE
SCOPUS
- Journal Title
- Journal of Clinical Microbiology
- Volume
- 58
- Number
- 1
- Start Page
- 1
- End Page
- 11
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/1393
- DOI
- 10.1128/JCM.01026-19
- ISSN
- 0095-1137
- Abstract
- Rapid and accurate detection of carbapenemase-producing Enterobacteriaceae (CPE) is critical for appropriate treatment and infection control. We compared a rapid fluorogenic assay using a carbapenem-based fluorogenic probe with other phenotypic assays: modified carbapenem inactivation method (mCIM), Carba NP test (CNP), and carbapenemase inhibition test (CIT). A total of 217 characterized isolates of Enterobacteriaceae were included as follows: 63 CPE; 48 non-carbapenemase-producing carbapenem-resistant Enterobacteriaceae (non-CP-CRE); 53 extended-spectrum beta-lactamase producers; and 53 third-generation-cephalosporin-susceptible isolates. The fluorogenic assay using bacterial colonies (Fluore-C) was conducted by lysing the isolates followed by centrifugation and mixing the supernatant with fluorogenic probe. In addition, for the fluorogenic assay using spiked blood culture bottles (Fluore-Direct), pellets were obtained via the saponin preparation method, which can directly identify the pathogens using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The fluorescence signal was measured over 50 min using a fluorometer. The fluorescent signal of CPE was significantly higher than that of non-CPE in both Fluore-C (median relative fluorescence units [RFU] [range], 5,814 [240 to 32,009] versus 804 [36 to 2,480], respectively; P < 0.0001) and Fluore-Direct (median RFU [range], 10,355 [1,689 to 31,463] versus 1,068 [428 to 2,155], respectively; P < 0.0001) tests. Overall, positive and negative percent agreements of Fluore-C, mCIM, CNP, CIT, and Fluore-Direct were 100% and 98.7%, 98.3% and 97.5%, 88.1% and 100%, 96.4% and 98.7%, and 98.3% and 98.1%, respectively. The relatively lower positive percent agreement (PPA) of CNP was mainly observed in OXA-type CPE. The fluorogenic assay showed excellent performance with bacterial colonies and also directly from positive blood cultures. We included many non-CP-CRE isolates for strict evaluation. The fluorogenic assay will be a useful tool for clinical microbiology laboratories.
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