Abstract

Increasing worldwide, prevalence of carbapenem-resistant gram-negative bacteria demands urgent a need for rapid detection and accurate identification of carbapenemases. The BD Phoenix CPO detect (PCD) assay possesses an in-built capacity for parallel susceptibility testing and detection of carbapenemases. Here, the ability of the assay to detect and classify carbapenemase production was tested in a collection of carbapenem-resistant Enterobacterales and non-fermentative gram-negative rods. The ability of the PCD assay to detect and classify carbapenemases was investigated in a collection of 194 clinical, carbapenem-resistant isolates (Enterobacterales [n = 65]; non-fermentative gram-negative rods [n = 129]). AST results were compared to MICS determined by gradient diffusion to determine accuracy of the PCD assay. The accuracy of the PCD assay to detect carbapenemases was compared to the results of molecular isolate characterization using a LDT multiplex carbapenemase PCR assay. All 194 isolates classified as carbapenem-resistant by reference susceptibility testing were also classified correctly as CRO by the PCD assay. Performance analysis of the PCD assay to detect carbapenemase production revealed an overall sensitivity of 98.29% and specificity of 17.95% for the detection of carbapenemase production. For the classification of carbapenemases classes A, B, and D, the PCD correctly classified 79.17% Enterobacterales and 67.16% non-fermentative gram-negative rods. The PCD assay is a reliable tool for the detection of carbapenem resistance and allows for parallel analysis of carbapenemase production. However, while sensitivity is high, low specificity in carbapenemase detection and erroneous classification demands mandatory confirmation by alternative methods, especially in non-fermentative gram-negative bacteria.

Highlights

  • Infections caused by carbapenem-resistant organisms (CRO) are emerging as a major challenge to public health [1, 2]

  • All 194 isolates classified as CRO by reference susceptibility testing (Vitek II, gradient strip) were found carbapenemresistant by the Phoenix CPO detect (PCD) assay

  • 65/ 79 (82.28 %) carbapenemase-negative isolates were identified as CPOs by the PCD

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Summary

Introduction

Infections caused by carbapenem-resistant organisms (CRO) are emerging as a major challenge to public health [1, 2]. Over the past couple of years, a marked increase in the prevalence of CRE has been reported worldwide [3], and infections caused by CRO are associated with a significant mortality [4]. The majority of these infections are associated with increased length of stay and mortality rates ranging from 30 to 70% [6]. Gram-negative organisms essentially become carbapenemresistant via two main routes, i.e., carbapenemase production or expression of cephalosporinases (AmpC) and/or extended spectrum β-lactamases (ESBL) in combination with cell wall permeability changes [7, 8]. Carbapenemases from molecular classes A, B, C, and D [9] are characterized by specific β-lactam hydrolytic profiles and susceptibility against inhibition by different β-lactamase inhibitors [10]

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