scholarly journals Performance of the BD Phoenix CPO detect assay for detection and classification of carbapenemase-producing organisms

Author(s):  
Laura Berneking ◽  
Anna Both ◽  
Benjamin Berinson ◽  
Armin Hoffmann ◽  
Marc Lütgehetmann ◽  
...  

AbstractIncreasing 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.

Author(s):  
Hari P. Nepal ◽  
Rama Paudel

Carbapenems are beta-lactam drugs that have broadest spectrum of activity. They are commonly used as the drugs of last resort to treat complicated bacterial infections. They bind to penicillin binding proteins (PBPs) and inhibit cell wall synthesis in bacteria. Important members that are in clinical use include doripenem, ertapenem, imipenem, and meropenem. Unlike other members, imipenem is hydrolyzed significantly by renal dehydropeptidase; therefore, it is administered together with an inhibitor of renal dehydropeptidase, cilastatin. Carbapenems are usually administered intravenously due to their low oral bioavailability. Most common side effects of these drugs include nausea, vomiting, diarrhea, skin rashes, and reactions at the infusion sites. Increasing resistance to these antibiotics is being reported throughout the world and is posing a threat to public health.  Primary mechanisms of carbapenem resistance include expulsion of drug and inactivation of the drug by production of carbapenemases which may not only hydrolyze carbapenem, but also cephalosporin, penicillin, and aztreonam. Resistance especially among Gram negative bacteria is of much concern since there are only limited therapeutic options available for infections caused by carbapenem resistant Gram-negative bacterial pathogens. Commonly used drugs to treat such infections include polymyxins, fosfomycin and tigecycline.


2021 ◽  
pp. 16-19
Author(s):  
N. I. Gabrielyan ◽  
V. G. Kormilitsyna ◽  
V. K. Zaletaeva ◽  
A. V. Krotevich ◽  
I. A. Miloserdov ◽  
...  

Detection of carbapenem resistance genes is a critical issue for hospitals due to possible recommendations for infection control and targeted therapy. The Cepheid Xpert instrument, a Carba-R test for the detection and differentiation of five common carbapenemase genes, was tested from September 2020 to February 2021. As part of the approbation, 20 tests were provided. This review presents the results of the approbation of a relatively regular sensitivity study on Siemens WalkAway‑96 plus. Cepheid Xpert Carba-R analysis has been shown to be an accurate and fast tool for detecting colonization by carbapenem-resistant gram-negative bacteria, which can help limit the spread of these organisms in hospitals.


2017 ◽  
Vol 63 (3) ◽  
pp. 723-730 ◽  
Author(s):  
Allison R McMullen ◽  
Melanie L Yarbrough ◽  
Meghan A Wallace ◽  
Angela Shupe ◽  
Carey-Ann D Burnham

Abstract BACKGROUND Carbapenemase-producing gram-negative bacteria (CP-GNB) are an urgent and expanding public health threat. Rapid and accurate identification of these organisms facilitates infection prevention efforts in healthcare facilities. The objective of our study was to evaluate methods to detect and identify CP-GNB. METHODS We examined 189 carbapenem-resistant GNB(CR-GNB), including Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii complex, using 3 different methods: 2 methods to screen isolates of GNB for carbapenemase production [the carbapenem inactivation method (CIM) and 2 chromogenic agars] and a molecular method (Cepheid GeneXpert Carba-R) to identify the mechanism of carbapenem resistance and the associated resistance genes (blaKPC, blaNDM, blaIMP, blaOXA-48-like, and blaVIM). RESULTS The CIM was a simple and inexpensive phenotypic screen to differentiate between CR-GNB and CP-GNB, with improved analytical performance characteristics and inter-reader correlation compared to the modified Hodge test. Both chromogenic agars evaluated (HardyCHROM CRE and chromID CARBA) were able to support growth of most of the organisms tested, including isolates possessing the blaOXA-48-like gene. However, these media had a low analytical specificity for carbapenemase production, with breakthrough of CR-GNB that did not produce a carbapenemase. The Xpert Carba-R assay was rapid and easy to perform, and demonstrated 100% positive and negative agreement for characterization of genetic determinants of carbapenem resistance. CONCLUSIONS Screening by CIM followed by the Xpert Carba-R PCR is an accurate method for detecting and characterizing CP-GNB, including Enterobacteriaceae, P. aeruginosa, and A. baumannii complex.


2021 ◽  
Vol 15 (7) ◽  
pp. 1807-1809
Author(s):  
Sadia Ikram ◽  
Anila Errum ◽  
Asma Inam ◽  
Farrukh Sarfaraz ◽  
Sadia Majeed ◽  
...  

Aim: To compare the resistance amongst Gram negative bacteria against imipenem and meropenem. Study Design: Prospective, non-randomized, descriptive study. Place and Duration of Study: Department of Microbiology, Mughal Laboratories, Lahore from 1stJuly 2019 to 31stDecember 2019. Methodology: One hundred culture samples received, bacteria isolated and their susceptibilities to imipenem and meropenem were compared. Organisms were recognized by the microbiological techniques according to the current standards and susceptibility testing was done according to the guidelines of Clinical and Laboratory Standards Institute (CLSI) 2020by using Kirby Bauer Disc diffusion method. Results: Salmonella typhi, Citrobacter species and Proteus species were 100% sensitive to imipenem. The rest of bacterial isolates had sensitivities to E. coli 88%, Acinetobacter 80%, Klebsiella species 67% and Peudomonas species 64%. The meropenem is highly resistant in all the bacteria as compared to imipenem. Conclusion: Increasing the trend of carbapenem resistance amongst Gram negative bacteria excluding Salmonella typhi was recorded. Key words: Gram negative rods, Resistance, Spectrum


2019 ◽  
Author(s):  
Prasanth Manohar ◽  
Murugavel Ragavi ◽  
Ashby Augustine ◽  
Hrishikesh MV ◽  
Nachimuthu Ramesh

AbstractBackgroundEmergence of carbapenem resistance mechanisms among Gram-negative bacteria is a worrisome health problem. Here, we focused on to identify the presence of carbapenem-resistant bacteria among the samples collected from hospital environments in Tamil Nadu.MethodsA total of 30 hospital environmental samples were collected between August 2017 and January 2018 from hospitals located in Chennai and Vellore such as lift switches, stair rails, switchboards, nursing desks, used nursing gloves, door handles, wheelchairs, touch screens, chairs and from pillars inside the hospitals.Results and discussionA total of 22 carbapenem-resistant Gram-negative bacteria were isolated that included Escherichia coli, Klebsiella sp., Enterobacter sp., Salmonella sp., Pseudomonas aeruginosa and Acinetobacter sp. Interestingly, blaGIM-1 was detected in Acinetobacter variabilis strain isolated in samples collected from hospitals. Unlike other studies, the identified GIM-1 was not plasmid encoded, and this is the first report for the presence of GIM-1 (German imipenemase) in India.ConclusionExtensive surveillance programs are necessary to trace the uncontrolled spread of carbapenem-resistance genes in order to reduce the rapid spread of resistance.


2018 ◽  
pp. 344-349
Author(s):  
Do Ogbolu ◽  
Ma Webber

Objective: To determine the role of extended-spectrum β-lactamases in carbapenem-resistant Gram-negative bacteria from south-western Nigeria. Methods: Twenty-seven carbapenem-resistant isolates that were found to be non-carbapenemase producers (15 Escherichia coli, 9 Klebsiella pneumoniae and 3 Pseudomonas aeruginosa) were further studied. These isolates were subjected to analysis including phenotypic and genotypic detection of various β-lactamases, efflux activity, outer membrane protein, plasmids replicon typing, detection of transferable genes and resistances and typing using random amplified polymorphic DNA tests. Results: No isolates demonstrated de-repression of efflux, but all showed either complete loss or reduced production of outer membrane proteins. Transconjugants from these strains contained various genes including plasmid-mediated quinolone resistance and extended-spectrum beta-lactamases. All the transconjugants carried the blaCTX-M-15 gene. The transconjugants had varying minimum inhibitory concentrations of carbapenems ranging from 0.03 μg/ml to 8 μg/ml. Varying resistances to other antimicrobial agents were also transferred with the plasmids. The donor isolates were not clonally related by molecular typing. Conclusion: Resistance to carbapenem antibiotics in this sample was not mediated only by carbapenemases but also by production of extended-spectrum β-lactamases (largely CTX-M-15), accompanied by protein loss. This was an important mechanism underpinning carbapenem resistance in these clinical isolates of various species.


10.3823/823 ◽  
2018 ◽  
Vol 8 (2) ◽  
Author(s):  
Monzer Hamze

Background. Antimicrobial resistance is a major public health problem worldwide. Numerous epidemiological studies reported that Lebanon is affected with high levels of antibiotic resistance. The aim of this study is to determine the prevalence and antibiotic susceptibility patterns of carbapenem resistant Gram negative bacteria in North Lebanon during the period 2015-2017. Methods. Carbapenem resistant Gram negative bacteria were isolated from patients referring to Nini hospital and Youssef hospital center. Identification and antibiotic susceptibility testing were performed through conventional tools according to the manufacturer’s recommended procedures and the recommendations of the European Committee on Antimicrobial Susceptibility Testing, respectively. Results. Overall, a total of 290 carbapenem resistant Gram negative bacteria were isolated. Escherichia coli was predominant and represented 39.3% of all isolates, followed by Pseudomonas aeruginosa (24.8%), Acinetobacter baumannii (22.8%), Klebsiella spp. (8.6%), Enterobacter spp. (6.6%), Pantoea spp. (1%), and Proteus vulgaris (0.3%). Our findings showed an alarming increase in the prevalence of carbapenem resistant bacteria every year. On the other hand, colistin, tigecycline, amikacin and fosfomycin remain the most effective agents against carbapenem resistant Gram negative bacteria. Conclusion. This study provided important new laboratory data that could support specialists in infectious diseases in North Lebanon to take the appropriate decision in the treatment of patients at risk for infections with carbapenem resistant Gram negative germs.


2017 ◽  
Vol 9 (04) ◽  
pp. 303-307 ◽  
Author(s):  
Shoorashetty Manohara Rudresh ◽  
Giriyapur Siddappa Ravi ◽  
Lakshminarayanappa Sunitha ◽  
Sadiya Noor Hajira ◽  
Ellappan Kalaiarasan ◽  
...  

Abstract PURPOSE: Detection of carbapenemases among Gram-negative bacteria (GNB) is important for both clinicians and infection control practitioners. The Clinical and Laboratory Standards Institute recommends Carba NP (CNP) as confirmatory test for carbapenemase production. The reagents required for CNP test are costly and hence the test cannot be performed on a routine basis. The present study evaluates modifications of CNP test for rapid detection of carbapenemases among GNB. MATERIALS AND METHODS: The GNB were screened for carbapenemase production using CNP, CarbAcineto NP (CANP), and modified CNP (mCNP) test. A multiplex polymerase chain reaction (PCR) was performed on all the carbapenem-resistant bacteria for carbapenemase genes. The results of three phenotypic tests were compared with PCR. RESULTS: A total of 765 gram negative bacteria were screened for carbapenem resistance. Carbapenem resistance was found in 144 GNB. The metallo-β-lactamases were most common carbapenemases followed by OXA-48-like enzymes. The CANP test was most sensitive (80.6%) for carbapenemases detection. The mCNP test was 62.1% sensitive for detection of carbapenemases. The mCNP, CNP, and CANP tests were equally sensitive (95%) for detection of NDM enzymes among Enterobacteriaceae. The mCNP test had poor sensitivity for detection of OXA-48-like enzymes. CONCLUSION: The mCNP test was rapid, cost-effective, and easily adoptable on routine basis. The early detection of carbapenemases using mCNP test will help in preventing the spread of multidrug-resistant organisms in the hospital settings.


2019 ◽  
Vol 69 (Supplement_7) ◽  
pp. S521-S528 ◽  
Author(s):  
Patrice Nordmann ◽  
Laurent Poirel

Abstract Carbapenem resistance in gram-negative bacteria has caused a global epidemic that continues to grow. Although carbapenemase-producing Enterobacteriaceae have received the most attention because resistance was first reported in these pathogens in the early 1990s, there is increased awareness of the impact of carbapenem-resistant nonfermenting gram-negative bacteria, such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. Moreover, evaluating the problem of carbapenem resistance requires the consideration of both carbapenemase-producing bacteria as well as bacteria with other carbapenem resistance mechanisms. Advances in rapid diagnostic tests to improve the detection of carbapenem resistance and the use of large, population-based datasets to capture a greater proportion of carbapenem-resistant organisms can help us gain a better understanding of this urgent threat and enable physicians to select the most appropriate antibiotics.


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