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Rapid Detection and Differentiation of KPC and MBL Carbapenemases among Enterobacterales Isolates by a Modified Combined-Disk Test.

ABSTRACT: This study was conducted to develop a cheap, rapid, and accurate modified combined-disk test (mCDT) approach to detect and differentiate KPC and MBL carbapenemases among clinical carbapenem-resistant Enterobacterales (CRE) isolates and simultaneously distinguish them from carbapenem-susceptible Enterobacterales (CSE) isolates. A total of 163 CRE and 90 third-generation cephalosporin-resistant Enterobacterales isolates were tested using imipenem and meropenem disks and different concentrations of carbapenemase inhibitors. The optimal sensitivity and specificity for detecting KPC carbapenemase were 97.2% and 100%, respectively. The sensitivity and specificity for detecting MBL carbapenemase were 100% and 100% with imipenem or meropenem and carbapenemase inhibitors within six hours. The inhibitory zone diameter of 18 mm for imipenem or meropenem disks without inhibitor could distinguish CRE from CSE isolates. Therefore, this mCDT approach may be a useful tool in clinical laboratories to detect CRE isolates and differentiate KPC and MBL producers, which is beneficial for patient management and hospital infection prevention and control.

SUBMITTER: Wei M

PROVIDER: S-EPMC8458999 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Rapid Detection and Characterization of Carbapenemases in Enterobacterales with a New Modified Carbapenem Inactivation Method, mCIMplus.

Project description:The worldwide emergence and spread of antimicrobial resistance in Gram-negative bacteria are severely limiting therapeutic options and thus constitute a major public health threat. The timely accurate detection of carbapenemase producers and the determination of carbapenemase class according to the Ambler classification can guide antimicrobial therapy and facilitate infection control measures. A modified version of the carbapenemase inactivation method (CIM), mCIM, was described and approved by the CLSI in 2017. We evaluated the performance of a faster new mCIM-based assay, mCIMplus, which can detect carbapenemase activity within 8 h and characterize the carbapenemase according to the Ambler classification in 20 h. A panel of 137 isolates producing carbapenemases (GES, IMP, KPC, NDM, OXA-48, OXA-48-like, and VIM enzymes) and 22 non-carbapenemase-producing isolates was used to evaluate the performance of mCIMplus. We evaluated the detection of carbapenemase activity at 8 and 20 h. Carbapenemase class was determined, with specific inhibitors, at 20 h. The sensitivities of mCIMplus were 99.3% at 8 h and 98.5% at 20 h. Its specificity was 100% regardless of culture time. Based on a decision algorithm, this test successfully identified the carbapenemase class for 98.4% of the tested isolates (127/129). Characterization was correct for 100, 95, and 100% of Ambler class A, B, and D isolates, respectively. This test can, therefore, be used to detect carbapenemase activity within 8 h and to determine carbapenemase class within 20 h. It constitutes a very affordable (<€1 per isolate) and reliable technique requiring only basic laboratory equipment.

| S-EPMC7587109 | biostudies-literature

NitroSpeed-Carba NP Test for Rapid Detection and Differentiation between Different Classes of Carbapenemases in Enterobacterales.

Project description:A biochemical test (NitroSpeed-Carba NP test) was developed to identify carbapenemase production in Enterobacterales and to discriminate between the different types of clinically significant carbapenemases (Ambler classes A, B, and D). It is based on two main features, namely, the hydrolysis by all β-lactamases, including carbapenemases of the nitrocefin substrate, and the capacity of ertapenem to prevent this hydrolysis for all β-lactamases except carbapenemases. Specific carbapenemase inhibitors of class A (avibactam, vaborbactam), class B (dipicolinic acid), and class D (avibactam) were used to inhibit the nitrocefin hydrolysis and to allow the identification of the carbapenemase types with a turnaround time of ca. 30 min. The test was evaluated with a collection of 248 clinical enterobacterial isolates, including 148 carbapenemase producers and 100 non-carbapenemase producers. Its overall sensitivity and specificity were 100% and 97%, respectively, including detection of all types of OXA-48-like carbapenemases. For the detection of the carbapenemase type, including strains that produce double carbapenemases, the sensitivity was 100%, 97%, and 100% for the detection of classes A, B, and D, respectively. This easy-to-implement test may contribute to optimization of the choice of the β-lactam/β-lactamase inhibitor combinations for treating infection due to carbapenemase producers.

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| S-EPMC4808189 | biostudies-literature

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| S-EPMC7470286 | biostudies-literature

Prospective evaluation of the OKN K-SeT assay, a new multiplex immunochromatographic test for the rapid detection of OXA-48-like, KPC and NDM carbapenemases.

Project description:ObjectivesThere is an urgent need for accurate and fast diagnostic tests capable of identifying carbapenemase producers. Here, we assessed the performance of a new multiplex lateral flow assay (OKN K -SeT) for the rapid detection of OXA-48-like, KPC and NDM carbapenemase-producing Enterobacteriaceae from culture colonies.MethodsTwo hundred collection isolates with characterized ?-lactamase content and 183 non-duplicate consecutive isolates referred to two National Reference Centres over a 2?month period in 2016 were used to evaluate the OKN K -SeT assay.ResultsThe assay correctly detected all 42 OXA-48-like-, 27 KPC- and 30 NDM-producing isolates from the collection panel, including 7 isolates that co-produced NDM and OXA-181 carbapenemases. No cross-reactivity was observed with non-targeted carbapenemases ( n? = ? 41) or with non-carbapenemase producers ( n? = ? 60). Prospectively, all OXA-48-like ( n? = ? 69), KPC ( n? = ? 9) and NDM ( n? = ? 19) carbapenemase-producing Enterobacteriaceae isolates were correctly detected, while 11 carbapenemase producers not targeted by the assay went undetected [VIM ( n? = ? 8) and OXA-23/OXA-58-like ( n? = ? 3)]. Overall, the sensitivity and specificity of the assay were 100%.ConclusionsThe OKN assay is efficient, rapid and easy to implement in the workflow of a clinical microbiology laboratory for the confirmation of OXA-48, NDM and KPC carbapenemases. This test represents a powerful diagnostic tool as it enables the rapid detection of the most clinically important carbapenemases without the need for more costly and less frequently available molecular assays.

| S-EPMC5890672 | biostudies-literature

Rapid Detection of bla<sub>KPC</sub> in Carbapenem-Resistant Enterobacterales Based on CRISPR/Cas13a.

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| S-EPMC10638124 | biostudies-literature

Evaluation of boronic acid disk tests for differentiating KPC-possessing Klebsiella pneumoniae isolates in the clinical laboratory.

Project description:The worldwide increase in the occurrence and dissemination of KPC beta-lactamases among gram-negative pathogens makes critical the early detection of these enzymes. Boronic acid disk tests using different antibiotic substrates were evaluated for detection of KPC-possessing Klebsiella pneumoniae isolates. A total of 57 genotypically confirmed KPC-possessing K. pneumoniae isolates with varying carbapenem MICs were examined. To measure the specificity of the tests, 106 non-KPC-possessing isolates (89 K. pneumoniae and 17 Escherichia coli isolates) were randomly selected among those exhibiting reduced susceptibility to cefoxitin, expanded-spectrum cephalosporins, or carbapenems. As many as 56, 53, and 40 of the non-KPC-possessing isolates harbored extended-spectrum beta-lactamases, metallo-beta-lactamases, and plasmid-mediated AmpC beta-lactamases, respectively. By use of CLSI methodology and disks containing imipenem, meropenem, or cefepime, either alone or in combination with 400 microg of boronic acid, all 57 KPC producers gave positive results (sensitivity, 100%) whereas all 106 non-KPC producers were negative (specificity, 100%). The meropenem duplicate disk with or without boronic acid demonstrated the largest differences in inhibition zone diameters between KPC producers and non-KPC producers. By use of disks containing ertapenem, all isolates were correctly differentiated except for five AmpC producers that gave false-positive results (sensitivity, 100%; specificity, 95.3%). These practical and simple boronic acid disk tests promise to be very helpful for the accurate differentiation of KPC-possessing K. pneumoniae isolates, even in regions where different broad-spectrum beta-lactamases are widespread.

| S-EPMC2643660 | biostudies-literature

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