Comparison of the Clinical Outcomes of Patients With Positive Xpert Carba-R Tests for Carbapenemase-Producing Enterobacterales According to Culture Positivity

Background We aimed to compare the clinical outcomes of patients with positive Xpert Carba-R assay results for carbapenemase-producing Enterobacterales (CPE) according to CPE culture positivity. Methods We retrospectively collected data for patients with positive CPE (positive Xpert Carba-R or culture) who underwent both tests from August 2018 to March 2021 in a 2700-bed tertiary referral hospital in Seoul, South Korea. We compared the clinical outcomes of patients positive for Xpert Carba-R according to whether they were positive (XPCP) or negative (XPCN) for CPE culture. Results Of 322 patients with CPE who underwent both Xpert Carba-R and culture, 313 (97%) were positive for Xpert Carba-R for CPE. Of these, 87 (28%) were XPCN, and 226 (72%) were XPCP. XPCN patients were less likely to have a history of previous antibiotic use (75.9% vs 90.3%; P = .001) and to have Klebsiella pneumoniae carbapenemase (21.8% vs 48.9%; P < .001). None of the XPCN patients developed infection from colonization within 6 months, whereas 13.4% (29/216) of the XPCP patients did (P < .001). XPCN patients had lower transmission rates than XPCP patients (3.0% [9/305] vs 6.3% [37/592]; P = .03). There was no significant difference in CPE clearance from positive culture results between XPCN and XPCP patients (40.0% [8/20] vs 26.7% [55/206]; P = .21). Conclusions Our study suggests that XPCN patients had lower rates of both infection and transmission than XPCP patients. The Xpert Carba-R assay is clinically useful not only for rapid identification of CPE but also for predicting risks of infection and transmission when performed along with culture.

Characterization of 2 Klebsiella pneumoniae carbapenemase–producing Enterobacterales isolated from canine rectal swabs

Globally, carbapenemase-producing Enterobacterales (CPE) cause life-threatening, hospital-acquired infections in people, and have been reported recently among veterinary patients. Organisms that produce a Klebsiella pneumoniae carbapenemase (KPC) are one of the most common CPE isolated from people but have been reported only rarely in animals. We characterized 2 KPC-producing Enterobacterales isolated from companion animal rectal swabs during the response to an outbreak caused by a strain of blaNDM-5 Escherichia coli. Both isolates were characterized by whole-genome sequencing (WGS) and analysis. The first isolate (case A) was from an immunosuppressed 6-y-old Yorkshire Terrier and was identified as E. coli (ST372) with a blaKPC-18 gene and an IncFII plasmid. The second isolate (case B) was from a 3-y-old Labrador Retriever with acute diarrhea and was identified as Citrobacter koseri with a blaKPC-2 gene, multiple plasmids (ColRNAI, pKPC-CAV1193), and a putative enterotoxin gene ( senB). Further research is needed to determine what role animals might play in the epidemiology of CPE in communities. It is imperative that all CPE isolated from companion animals be fully characterized by WGS and the associated case examined. All veterinary isolates should be sequenced and shared for surveillance, monitoring, and investigation purposes.

Carbapenemase Producing Klebsiella pneumoniae (KPC): What Is the Best MALDI-TOF MS Detection Method

Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria is a group of highly dangerous antibiotic resistant Gram-negative Enterobacteriaceae. They cause infections associated with significant morbidity and mortality. Therefore, the rapid detection of KPC-producing bacteria plays a key role in clinical microbiology. Matrix assisted laser desorption/ionization time-of- flight (MALDI-TOF) is a rapidly evolving technology that finds application in various clinical, scientific, and industrial disciplines. In the present study, we demonstrated three different procedures of carbapenemase-producing K. pneumoniae (KPC) detection. The most basic model of MALDI-TOF instrument MS Microflex LT was used, operating in the linear ion-positive mode, commonly used in modern clinical laboratories. The first procedure was based on indirect monitoring of carbapenemase production with direct detection of hydrolyzed carbapenem antibiotic degradation products in the mass spectrum. The second procedure was based on direct detection of blaKPC accompanying peak with an 11,109 Da in the mass spectrum of carbapenemase-producing K. pneumoniae (KPC), which represents the cleaved protein (pKpQIL_p019) expressed by pKpQIL plasmid. In addition, several unique peaks were detected in the carbapenemase-producing K. pneumoniae (KPC) mass spectrum. The third procedure was the identification of carbapenemase-producing K. pneumoniae (KPC) based on the protein fingerprint using local database created from the whole mass spectra. By comparing detection procedures, we determined that the third procedure was very fast and relatively easy. However, it requires previous verification of carbapenemase-producing K. pneumoniae (KPC) using other methods as genetic blaKPC identification, detection of carbapenem degradation products, and accompanying peak with 11,109 Da, which represents cleaved pKpQIL_p019 protein expressed by pKpQIL plasmid. Detection of carbapenemase-producing K. pneumoniae using MALDI-TOF provides fast and accurate results that may help to reduce morbidity and mortality in hospital setting when applied in diagnostic situations.

ASSISTÊNCIA DE ENFERMAGEM A PACIENTES COLONIZADOS POR KLEBSIELLA PNEUMONIAE CARBAPENEMASE (KPC)

Introdução: As Infecções relacionadas à assistência à saúde (IRAS) são doenças adquiridas durante a prestação de cuidados na saúde, acometendo o paciente após a hospitalização, nos últimos anos as IRAS elevaram-se em nível mundial, dentre elas, podemos citar o elevado índice de infecções causadas por cepas resistentes de Klebsiella pneumoniae carbapenemase (KPC), produtoras de carbapenemase. Nesse sentido, destaca-se que a enfermagem, por atuar ininterruptamente na assistência direta ao usuário realizando procedimentos invasivos e potencialmente contaminados, tem responsabilidade na profilaxia e no controle das IRAS. Objetivo: Analisar a assistência de enfermagem na prevenção e controle de infecções causadas por Klebsiella pneumoniae carbapenemase. Material e Método: Trata-se de uma revisão bibliográfica reflexiva, descritiva de cunho qualitativa, realizada através da base de dados Google Acadêmico. Para isso, utilizou-se os seguintes descritores em português: assistência de enfermagem, Klebsiella pneumoniae, Controle de Infecções e Segurança do Paciente. Houve restrição temporal entre os anos de 2017 a 2021. Resultados: Observaram-se os desafios para a prevenção e controle das infecções por KPC, no qual se destacam a resistência bacteriana aos antimicrobianos, o processamento de produtos para saúde e o comportamento dos profissionais de saúde diante dos protocolos de infecção hospitalar. Dessa forma, salienta-se que o enfermeiro não só dentro da comissão de controle de infecção hospitalar (CCIH), que é responsável pela elaboração, programação e desenvolvimento de ações de vigilância epidemiológica das infecções hospitalares, educação e treinamento das equipes e controle do uso racional de antimicrobianos, germicidas e materiais médico-hospitalares, mas de maneira geral, atua na supervisão e execução de medidas que alcancem índices positivos para uma assistência segura, com intuito de minimizar a disseminação da KPC. Conclusão: Portanto, evidencia-se a extensa atuação da enfermagem, de maneira geral, afirmando a importância da adesão de diferentes estratégias e a continuidade das já existentes, como: os hospitais devem desenvolver a capacitação dos profissionais; a equipe multiprofissional deve seguir o protocolo de medidas profiláticas, no qual, a instituição deve disponibilizar os recursos necessários; vigilância contínua da CCIH, entre outros métodos para minimizar a disseminação de patógenos e garantir o cuidado ao paciente.

Clonal Spread and Intra- and Inter-Species Plasmid Dissemination Associated With Klebsiella pneumoniae Carbapenemase-Producing Enterobacterales During a Hospital Outbreak in Barcelona, Spain

Objectives: The study aimed to characterize the clonal spread of resistant bacteria and dissemination of resistance plasmids among carbapenem-resistant Enterobacterales at a tertiary hospital in Catalonia, Spain.Methods: Isolates were recovered from surveillance rectal swabs and diagnostic samples. Species identification was by matrix-assisted laser desorption ionization-time time of flight mass spectrometry (MALDI-TOF MS). Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Antimicrobial susceptibility was assessed by gradient-diffusion and carriage of bla genes was detected by PCR. Plasmid typing, conjugation assays, S1-PFGE studies and long-read sequencing were used to characterize resistance plasmids.Results: From July 2018 to February 2019, 125 Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacterales were recovered from 101 inpatients from surveillance (74.4%) or clinical samples (25.6%), in a tertiary hospital in Barcelona. Clonality studies identified a major clone of Klebsiella pneumoniae belonging to sequence type ST15 and additional isolates of K. pneumoniae, Escherichia coli and Enterobacter sp. from different STs. All isolates but one carried the blaKPC–2 allelic variant. The blaKPC–2 gene was located in an IncFIIk plasmid of circa 106 Kb in a non-classical Tn4401 element designated NTEKPC-pMC-2-1. Whole-genome sequencing revealed different rearrangements of the 106 Kb plasmid while the NTEKPC-pMC-2-1 module was highly conserved.Conclusion: We report a hospital outbreak caused by the clonal dissemination of KPC-producing ST15 K. pneumoniae but also the intra- and inter-species transmission of the blaKPC–2 gene associated with plasmid conjugation and/or transposon dissemination. To our knowledge, this is the first report of an outbreak caused by KPC-producing Enterobacterales isolated from human patients in Catalonia and highlights the relevance of surveillance studies in the early detection and control of antibiotic resistant high-risk clones.

Phenotypic and molecular evaluation of biofilm formation in Klebsiella pneumoniae carbapenemase (KPC) isolates obtained from a hospital of Pelotas, RS, Brazil

Introduction. A significant cause of mortality in the intensive care unit (ICU) is multidrug-resistant (MDR) Gram-negative bacteria, such as Klebsiella pneumoniae carbapenemase (KPC). Biofilm production is a key factor in KPC colonization and persistence in the host, making the treatment difficult. Gap Statement. The aim of this study was to evaluate the antibiotic resistance, molecular and phenotypic biofilm profiles of 12 KPC isolates associated with nosocomial infection in a hospital in Pelotas, Rio Grande do Sul, Brazil. Methodology. Clinical isolates were obtained from different sources, identified and characterized by antibiotic resistance and carbapenemase synthesis following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Polymerase chain reaction (PCR) was used to evaluate the presence of carbapenemase (blaKPC ) and biofilm formation-associated genes (fimA, fimH, rmpA, ecpA, mrkD and wabG). Additionally, phenotypic evaluation of in vitro biofilm formation capacity was evaluated by Congo red agar (CRA) assay and the crystal violet staining method. Results. The 12 isolates evaluated in this study presented the blaKPC gene and were positive for synthesizing carbapenemases in vitro. In the carbapenem class, 83.3 % isolates were resistant and 16.7 % intermediately resistant to imipenem and meropenem. Molecular analyses found that the fimA and wabG genes were detected in 75 % of isolates, while fimH and ecpA were detected in 42 % and mrkD were detected in 8.3 % (1). The CRA assay demonstrated that all isolates were slime producers and 91.7 % (11) of isolates were classified as strong and 8.3 % (1) as moderate biofilm producers by the crystal violet staining method. The optical density (OD540nm) for strong biofilm formers ranged from 0.80±0.05 to 2.47±0.28 and was 0.55±0.12 for moderate biofilm formers. Conclusion. Our study revealed a high level of antibiotic resistance and biofilm formation in KPC isolates obtained from a hospital in Pelotas, RS, Brazil.

Treatment of UTIs Due to Klebsiella pneumoniae Carbapenemase-Producers: How to Use New Antibiotic Drugs? A Narrative Review

Background: K. pneumoniae is one of the bacteria most frequently causing health care-associated urinary tract infections, and increasingly incriminating Klebsiella pneumoniae carbapenemase producers (KPCp). Most infections caused by KPCp are nosocomial and might cause serious issues, even leading to death in half of the reported cases. Our aim was to identify the best strategy, based on available scientific data, for the use of new antibiotic treatments to manage KPCp UTIs. Methods: this narrative review of the literature was performed according to the criteria of preferred reporting items for systematic review and meta-analyses statement (PRISMA) (2020). Results and Conclusions: KPCp-UTIs are a real challenge for physicians. While cefiderocol, meropenem-vaborbactam, ceftazidim-avibactam, and imipenem-relebactam represent a major step forward in the treatment of these UTIs, no guidelines are currently available, in view of choosing the most appropriate treatment, in each specific case.

1268. In Vitro Activity of Ceftazidime-Avibactam and Comparators against KPC-Producing Enterobacterales and Pseudomonas aeruginosa Collected in China as Part of the ATLAS Global Surveillance Program in 2019

Background Among Gram-negative bacteria, the rapid spread of carbapenemases has limited therapeutic options. Klebsiella pneumoniae carbapenemase (KPC), an Ambler class A serine β-lactamase, presents a particular challenge as it has become widespread, first identified in an isolate collected in the United States and thereafter moving throughout the world, including China. Fortunately, the β-lactamase inhibitor avibactam is a potent inhibitor of KPC, rendering many Enterobacterales and some P. aeruginosa isolates that carry KPC susceptible to ceftazidime-avibactam (CAZ-AVI) in vitro. This study reports on the in vitro activity of CAZ-AVI and comparators against Enterobacterales and P. aeruginosa isolates collected in China as part of the Antimicrobial Testing Leadership and Surveillance (ATLAS) program in 2019. Methods 1,443 non-duplicate Enterobacterales and 522 P. aeruginosa isolates were collected from 17 clinical sites in China in 2019. Susceptibility testing was done using broth microdilution according to CLSI guidelines and interpreted using CLSI 2021 breakpoints. 143/177 meropenem non-susceptible Enterobacterales isolates and 150/187 meropenem non-susceptible P. aeruginosa isolates were interrogated by whole genome sequencing (WGS; Illumina 2x150 bp reads). Results Enterobacterales isolates exhibited higher % susceptibility (% S) to CAZ-AVI than all comparators tested (96.0% S; Table). The addition of AVI to CAZ resulted in an increase in susceptibility from 61.3% to 96.0% in the overall collection of Enterobacterales isolates. 96.0% of KPC-positive Enterobacterales, and 67.8% of the meropenem non-susceptible sub-population were susceptible to CAZ-AVI, against which comparators were less active (≤42.9 % S). Among P. aeruginosa isolates, 89.8% were susceptible to CAZ-AVI, more than for any comparator except amikacin (AMK; 94.4% S). Against meropenem non-susceptible and KPC-carrying P. aeruginosa sub-populations more were susceptible to CAZ-AVI (75.9% and 83.3% S, respectively) and AMK (87.2% and 100% S, respectively) than to other comparators (≤40.6% and ≤8.3% S, respectively). Results Table Conclusion CAZ-AVI demonstrated very good in vitro activity against Enterobacterales and P. aeruginosa isolates from China, including those that harbor KPC. Disclosures Mark G G. Wise, PhD, IHMA (Employee)Pfizer, Inc. (Independent Contractor) Krystyna Kazmierczak, PhD, IHMA (Employee)Pfizer, Inc. (Independent Contractor) Gregory Stone, PhD, AztraZeneca (Shareholder, Former Employee)Pfizer, Inc. (Employee) Daniel F. Sahm, PhD, IHMA (Employee)Pfizer, Inc. (Independent Contractor)