Investigation of carbapenemase and mcr-1 genes in carbapenem-resistant Klebsiella pneumoniae isolates

Introduction: Carbapenem-resistant Klebsiella pneumoniae are a major problem. We aimed to investigate carbapenemase-encoding genes and transferable mcr-1 genes among 57 carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates from hospitalized patients. Methodology: Antibiotic susceptibility tests were performed by Phoenix (BD). Results for ertapenem and colistin were confirmed by gradient diffusion and microdilution methods. Carbapenemase and mcr-1 genes were investigated by Polymerase Chain Reaction (PCR). Results: Thirty-two (56.14%) isolates were from intensive care units (ICU). Antibiotic resistance rates by Phoenix: 52.63% for amikacin; 73.69% trimethoprim sulfamethoxazole; 91.23% cefepime; 82.46% tigecycline; 59.65% colistin. Carbapenemases positivity: 82.45% (47) for blaOXA-48, 40.35% (23) blaOXA-55, 3.50% (2) blaOXA-51, 1.75% (1) blaOXA-23, 1.75% (1) blaOXA-24, 1.75% (1) blaIMP. blaOXA-58, blaKPC, blaNDM-1, and blaVIM were not detected. Twenty (35.08%) isolates had both blaOXA-48 and blaOXA-55. Three isolates were mcr-1 (+) and blaOXA-48 (+). One mcr-1 (+) isolates was blaOXA-51 (+). One colistin sensitive isolate determined by Phoenix, was found colistin resistant by microdilution. Conclusion: OXA-48 and OXA-55 co-harboring isolates and mcr-1 gene (+) isolates were spreading. Automated colistin susceptibility results should be confirmed by microdilution method. Resistance mechanisms in Enterobacteriaceae should be determined and the isolates should be monitored by molecular epidemiological methods. Effective infection control measures will contribute to reduce risk of antibiotic resistant bacterial infections and dissemination of antibiotic resistance.

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Molecular Epidemiology and Characterization of Carbapenem-Resistant Klebsiella pneumoniae Isolated from Urine at a Teaching Hospital in Taiwan

Microorganisms ◽

10.3390/microorganisms9020271 ◽

2021 ◽

Vol 9 (2) ◽

pp. 271

Author(s):

Yuarn-Jang Lee ◽

Chih-Hung Huang ◽

Noor Andryan Ilsan ◽

I-Hui Lee ◽

Tzu-Wen Huang

Keyword(s):

Klebsiella Pneumoniae ◽

Teaching Hospital ◽

Efflux Pump ◽

Resistance Mechanisms ◽

Pcr Analysis ◽

Carbapenem Resistant ◽

High Prevalence ◽

Antimicrobial Susceptibility Tests ◽

Susceptibility Tests ◽

Tract Infections

Urinary tract infections (UTIs) are common in clinics and hospitals and are associated with a high economic burden. Enterobacterium Klebsiella pneumoniae is a prevalent agent causing UTIs. A high prevalence of carbapenem-resistant K. pneumoniae (CRKP) has emerged recently and is continuing to increase. Seventeen urinary CRKP isolates collected at a teaching hospital in Taiwan from December 2016 to September 2017 were analyzed to elucidate their drug resistance mechanisms. Two-thirds of the isolates were obtained from outpatients. Antimicrobial susceptibility tests demonstrated multidrug resistance in all the isolates. Multilocus sequence typing analysis showed high diversity among the isolates. PCR analysis demonstrated the presence of carbapenemases in three isolates. All isolates carried at least one other extended-spectrum β-lactamase, including TEM, DHA, and CTX-M. Fifteen isolates contained mutations in one of the outer membrane porins that were assessed. The expression levels of the acrB and/or oqxB efflux pump genes, as determined by qRT-PCR, were upregulated in 11 isolates. Six isolates might have utilized other efflux pumps or antimicrobial resistance mechanisms. These analyses demonstrated a highly diverse population and the presence of complex resistance mechanisms in urinary isolates of K. pneumoniae.

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Spread of Carbapenemase Producing Klebsiella pneumoniae Isolates in Our Hospital: Investigation of Molecular Typing and Clonal Relationship

Türk Mikrobiyoloji Cemiyeti Dergisi ◽

10.5222/tmcd.2021.09226 ◽

2021 ◽

Author(s):

Reyhan Kiş ◽

Ebru Demiray Gündüz ◽

Ayşe Nur Sarı ◽

Zeynep Gülay

Keyword(s):

Intensive Care ◽

Klebsiella Pneumoniae ◽

Carbapenem Resistance ◽

Hospital Staff ◽

Control Measures ◽

Automated System ◽

Microbiology Laboratory ◽

Clonal Relationship ◽

Infection Control Measures ◽

Carbapenem Resistant

Objective: Carbapenem resistance has been reported with increasing frequency among members of Enterobacterales, especially in the last 10 years. Screening and detection of carbapenemase-producing isolates is important in terms of both directing the treatment and preventing its spread. In our study, it was aimed to determine the carbapenemase types and molecular epidemiological relationships of carbapenem resistant Klebsiella pneumoniae isolates, which were isolated sequentially from the samples sent to microbiology laboratory of our hospital. Method: A total of 32 carbapenem-resistant K. pneumoniae isolates of the samples sent to microbiology laboratory between July and September 2014, were included in the study. In addition to classical methods, identification of isolates at species level was made with BD Phoenix ID/AST automated system. Carbapenemase types (blaOXA-48, blaNDM, blaIMP, blaKPC, blaVIM and blaGES) of the isolates were investigated by PCR. The clonal relationship between the isolates was assessed with PFGE. Results: It was noted that 18 isolates were obtained from intensive care units, 9 from inpatient and 5 from outpatient departments. The blaOXA48 gene was found in all isolates while the other carbapenemase genes were not found. It was determined that strains were isolated from 32 patients in our hospital had 12 different PFGE pulsotypes, named as A-L. Among these, the most common ones were B (n=18) and closely related B1 pattern (n=2). The remaining isolates were represented by 11 different types. It was observed that the first isolate with B pulsotype was responsible for the spread of the outbreak from General Intensive Care Unit. Conclusion: It has been thought that the spread of carbapenem- resistant K. pneumoniae isolates in the hospital was probably occurred through the transfer of isolates from patients with gastrointestinal colonization to other patients through hospital staff. Therefore, the spread of the isolates in hospitals can be limited by detecting colonization with active surveillance programs and by applying contact isolation and effective infection control measures.

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The epidemiology, risk factors and outcomes of Carbapenem-resistant Klebsiella pneumoniae infections in the intensive care unit from 2012 to 2018

10.21203/rs.3.rs-17412/v1 ◽

2020 ◽

Author(s):

Ping Wang ◽

Xiaocui Zou ◽

Boting Zhou ◽

Tao Yin

Keyword(s):

Intensive Care Unit ◽

Intensive Care ◽

Klebsiella Pneumoniae ◽

Control Measures ◽

Icu Patients ◽

Infection Control Measures ◽

Carbapenem Resistant ◽

Incidence And Mortality ◽

Changing Trend ◽

Different Sources

Abstract Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an increasing globally threat for human health, but the trends and clinical characteristics of CRKP infections in the intensive care unit(ICU) remain uninvestigated.Methods: A retrospective study was conducted among ICU patients infected with KP isolates from January 2012 to December 2018. Carbapenem resistant to Klebsiella pneumoniae was defined according to Clinical and Laboratory Standards Institute (CLSI) criteria. The incidence and changing trend of CRKP were determined. CRKP patient sources, specimen types, infection sources and outcomes were investigated. Results: There were 256(40.13%) patients with CRKP and 382(59.87%) patients with CSKP. The incidence of CRKP increased from 2012(11.11%) to 2017(63.48%) and decreased in 2018(51.52%). The proportion of isolates not susceptible to three carbapenems increased from 0 to 98.04%. The rates of CRKP isolated from blood, wound, urine and pleural fluid were higher than that of CSKP. CRKP infections were mainly ICU acquired, rather than input acquired. Conclusion: The incidence of CRKP was high in ICU, but showed a downward trend. Implementation of different infection control measures to different sources of patients, specimen types, and KP infections are necessary. Surveillance data will be needed for ICU patients to decrease the incidence and mortality of CRKP.

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Post-antibiotic era in hemodialysis? Two case reports of simultaneous colonization and bacteremia by multidrug-resistant bacteria

Brazilian Journal of Nephrology ◽

10.1590/2175-8239-jbn-2020-0070 ◽

2020 ◽

Author(s):

Johanna M. Vanegas ◽

Lorena Salazar-Ospina ◽

Gustavo A. Roncancio ◽

Julián Builes ◽

Judy Natalia Jiménez

Keyword(s):

Pseudomonas Aeruginosa ◽

Bacterial Infections ◽

Case Reports ◽

Multidrug Resistant ◽

Resistance Mechanisms ◽

Control Measures ◽

Resistant Bacteria ◽

Multidrug Resistant Bacteria ◽

E Coli ◽

Carbapenem Resistant

ABSTRACT The emergence of resistance mechanisms not only limits the therapeutic options for common bacterial infections but also worsens the prognosis in patients who have conditions that increase the risk of bacterial infections. Thus, the effectiveness of important medical advances that seek to improve the quality of life of patients with chronic diseases is threatened. We report the simultaneous colonization and bacteremia by multidrug-resistant bacteria in two hemodialysis patients. The first patient was colonized by carbapenem- and colistin-resistant Klebsiella pneumoniae, carbapenem-resistant Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus (MRSA). The patient had a bacteremia by MRSA, and molecular typing methods confirmed the colonizing isolate was the same strain that caused infection. The second case is of a patient colonized by extended-spectrum beta-lactamases (ESBL)-producing Escherichia coli and carbapenem-resistant Pseudomonas aeruginosa. During the follow-up period, the patient presented three episodes of bacteremia, one of these caused by ESBL-producing E. coli. Molecular methods confirmed colonization by the same clone of ESBL-producing E. coli at two time points, but with a different genetic pattern to the strain isolated from the blood culture. Colonization by multidrug-resistant bacteria allows not only the spread of these microorganisms, but also increases the subsequent risk of infections with limited treatments options. In addition to infection control measures, it is important to establish policies for the prudent use of antibiotics in dialysis units.

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Molecular Epidemiology of Carbapenem-Resistant Klebsiella pneumoniae Infections in Southwest China

10.21203/rs.3.rs-34601/v1 ◽

2020 ◽

Author(s):

Xiaopin Hu ◽

Guohang Yuan ◽

Yaoyao Wu ◽

Weijia Liu ◽

Xiangyan Zhang ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Southwest China ◽

Antibiotic Use ◽

Resistance Mechanisms ◽

Control Measures ◽

Invasive Procedures ◽

Carbapenem Resistant ◽

Tertiary Hospitals ◽

Resistance Rates ◽

Epidemiological Characteristics

Abstract Background: We determined epidemiological characteristics and resistance mechanisms of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains found in Southwest China and assessed disease burden to provide evidence-based strategies for control and treatment of CRKP infection. Methods: A total of 159 strains of CRKP were isolated from sputa, blood, urine, ascites and wound secretions from three tertiary hospitals in Southwest China between August 1st, 2018 and December 31st, 2019. The sensitivity of each strain to 12 antibiotic agents was determined by micro-broth dilution. Identification of carbapenemase genes and multi-locus sequence typing (MLST) were performed using polymerase chain reaction (PCR). The disease burdens of patients with CRKP were assessed based on invasive procedures, antibiotic use, laboratory tests and clinical outcomes. Results: Of 159 CRKP strains analyzed, 50.9% were isolated from sputum samples. The percentage of patients who underwent invasive procedures before positive cultures for CRKP were detected was 96.3%. The mortality of blood infection was highest (66.6%) among patients with CRKP infection. All strains were insensitive to carbapenems. The resistance rates to levofloxacin and amikacin were 85.5% and 81.8%, respectively. All CRKP strains produced carbapenemases, with a majority of isolates (81.1%) producing KPC-2. The MICs of strains harbouring both KPC-2 and NDM-1 were higher than those of strains with only KPC-2 or NDM-1. ST11 is the most popular clonotype found in Southwest China. Conclusions: CRKP strains in Southwest China are characterized by strong drug resistance and associated with poor clinical prognoses. It is therefore urgent to both strengthen control measures and improve prevention awareness.

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Antimicrobial Stewardship Program, COVID-19, and Infection Control: Spread of Carbapenem-Resistant Klebsiella Pneumoniae Colonization in ICU COVID-19 Patients. What Did Not Work?

Journal of Clinical Medicine ◽

10.3390/jcm9092744 ◽

2020 ◽

Vol 9 (9) ◽

pp. 2744 ◽

Cited By ~ 4

Author(s):

Beatrice Tiri ◽

Emanuela Sensi ◽

Viola Marsiliani ◽

Mizar Cantarini ◽

Giulia Priante ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Infection Control ◽

Prone Position ◽

Antimicrobial Stewardship ◽

Control Measures ◽

Resistant Bacteria ◽

Infection Control Measures ◽

Carbapenem Resistant ◽

Stewardship Program ◽

Prolonged Contact

The Italian burden of disease associated with infections due to antibiotic-resistant bacteria has been very high, largely attributed to Carbapenem-Resistant Klebsiella pneumoniae (CR-Kp). The implementation of infection control measures and antimicrobial stewardship programs (ASP) has been shown to reduce healthcare-related infections caused by multidrug resistance (MDR) germs. Since 2016, in our teaching hospital of Terni, an ASP has been implemented in an intensive care unit (ICU) setting, with the “daily-ICU round strategy” and particular attention to infection control measures. We performed active surveillance for search patients colonized by Carbapenem-Resistant Enterobacteriaceae (CRE). In March 2020, coronavirus disease 2019 (COVID-19) arrived and the same ICU was reserved only for COVID-19 patients. In our retrospective observational study, we analyzed the bimonthly incidence of CRE colonization patients and the incidence of CRE acquisition in our ICU during the period of January 2019 to June 2020. In consideration of the great attention and training of all staff on infection control measures in the COVID-19 era, we would have expected a clear reduction in CRE acquisition, but this did not happen. In fact, the incidence of CRE acquisition went from 6.7% in 2019 to 50% in March–April 2020. We noted that 67% of patients that had been changed in posture with prone position were colonized by CRE, while only 37% of patients that had not been changed in posture were colonized by CRE. In our opinion, the high intensity of care, the prone position requiring 4–5 healthcare workers (HCWs), equipped with personal protective equipment (PPE) in a high risk area, with extended and prolonged contact with the patient, and the presence of 32 new HCWs from other departments and without work experience in the ICU setting, contributed to the spread of CR-Kp in our ICU, determining an increase in CRE acquisition colonization.

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A Prolonged Outbreak of KPC-3-Producing Enterobacter cloacae and Klebsiella pneumoniae Driven by Multiple Mechanisms of Resistance Transmission at a Large Academic Burn Center

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01516-16 ◽

2016 ◽

Vol 61 (2) ◽

Cited By ~ 32

Author(s):

Hajime Kanamori ◽

Christian M. Parobek ◽

Jonathan J. Juliano ◽

David van Duin ◽

Bruce A. Cairns ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Molecular Mechanisms ◽

Enterobacter Cloacae ◽

The United States ◽

Multidrug Resistant ◽

Control Measures ◽

Content Type ◽

Burn Center ◽

Infection Control Measures ◽

Carbapenem Resistant

ABSTRACT Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacter cloacae has been recently recognized in the United States. Whole-genome sequencing (WGS) has become a useful tool for analysis of outbreaks and for determining transmission networks of multidrug-resistant organisms in health care settings, including carbapenem-resistant Enterobacteriaceae (CRE). We experienced a prolonged outbreak of CRE E. cloacae and K. pneumoniae over a 3-year period at a large academic burn center despite rigorous infection control measures. To understand the molecular mechanisms that sustained this outbreak, we investigated the CRE outbreak isolates by using WGS. Twenty-two clinical isolates of CRE, including E. cloacae (n = 15) and K. pneumoniae (n = 7), were sequenced and analyzed genetically. WGS revealed that this outbreak, which seemed epidemiologically unlinked, was in fact genetically linked over a prolonged period. Multiple mechanisms were found to account for the ongoing outbreak of KPC-3-producing E. cloacae and K. pneumoniae. This outbreak was primarily maintained by a clonal expansion of E. cloacae sequence type 114 (ST114) with distribution of multiple resistance determinants. Plasmid and transposon analyses suggested that the majority of bla KPC-3 was transmitted via an identical Tn4401b element on part of a common plasmid. WGS analysis demonstrated complex transmission dynamics within the burn center at levels of the strain and/or plasmid in association with a transposon, highlighting the versatility of KPC-producing Enterobacteriaceae in their ability to utilize multiple modes to resistance gene propagation.

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Molecular Analysis of Ciprofloxacin Resistance Mechanisms in Malaysian ESBL-ProducingKlebsiella pneumoniaeIsolates and Development of Mismatch Amplification Mutation Assays (MAMA) for Rapid Detection ofgyrAandparCMutations

BioMed Research International ◽

10.1155/2014/601630 ◽

2014 ◽

Vol 2014 ◽

pp. 1-10 ◽

Cited By ~ 15

Author(s):

Farah Al-Marzooq ◽

Mohd Yasim Mohd Yusof ◽

Sun Tee Tay

Keyword(s):

Klebsiella Pneumoniae ◽

Infection Control ◽

Molecular Analysis ◽

Rapid Detection ◽

Resistance Mechanisms ◽

Control Measures ◽

Quinolone Resistance ◽

Infection Control Measures ◽

High Prevalence ◽

Ciprofloxacin Resistance

Ninety-three Malaysian extended-spectrumβ-lactamase (ESBL)-producingKlebsiella pneumoniaeisolates were investigated for ciprofloxacin resistance. Two mismatch amplification mutation (MAMA) assays were developed and used to facilitate rapid detection ofgyrAandparCmutations. The isolates were also screened for plasmid-mediated quinolone resistance (PMQR) genes includingaac(6′)-Ib-cr, qepA, andqnr. Ciprofloxacin resistance (MICs4–≥32 μg/mL) was noted in 34 (37%) isolates, of which 33 isolates had multiple mutations either ingyrAalone(n=1)or in bothgyrAandparCregions(n=32).aac(6′)-Ib-crwas the most common PMQR gene detected in this study(n=61), followed byqnrBandqnrS(n=55and 1, resp.). Low-level ciprofloxacin resistance (MICs 1-2 μg/mL) was noted in 40 (43%) isolates carryingqnrBaccompanied by eitheraac(6′)-Ib-cr(n=34)or a singlegyrA83 mutation(n=6). Ciprofloxacin resistance was significantly associated with the presence of multiple mutations ingyrAandparCregions. While the isolates harbouringgyrAand/orparCalteration were distributed into 11 PFGE clusters, no specific clusters were associated with isolates carrying PMQR genes. The high prevalence of ciprofloxacin resistance amongst the Malaysian ESBL-producingK. pneumoniaeisolates suggests the need for more effective infection control measures to limit the spread of these resistant organisms in the hospital.

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Molecular characteristics of antibiotic resistance Escherichia coli and Klebsiella pneumoniae strains isolated from hospitalized patients in Tehran, Iran

10.21203/rs.3.rs-120490/v1 ◽

2020 ◽

Author(s):

Javad Yasbolaghi Sharahi ◽

Ali Hashemi ◽

Abdollah Ardebili ◽

Sara Davoudabadi

Keyword(s):

Resistance Genes ◽

Large Scale ◽

Antimicrobial Agents ◽

Resistance Mechanisms ◽

Control Measures ◽

Molecular Characteristics ◽

Colistin Resistance ◽

E Coli ◽

Infection Control Measures ◽

Microdilution Method

Abstract Background We evaluated the distribution of carbapenem and colistin resistance mechanisms of clinical E. coli and K. pneumoniae isolates from Iran. Methods Antibiotic susceptibility testing was performed by disc diffusion and microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) guideline. The presence of extended spectrum β-lactamases (ESBLs) genes, carbapenemase genes, as well as fosfomycin resistance genes, and colistin resistance genes was also examined by PCR-sequencing. The ability of biofilm formation was assessed with crystal violet staining method. The expression colistin resistance genes were measured by quantitative reverse transcription-PCR (RT-qPCR) analysis to evaluate the association between gene upregulation and colistin resistance. Genotyping was performed using the multi-locus sequencing typing (MLST). Results From 2016 to 2018, a total of 165 clinical isolates of K. pneumoniae and E. coli were collected. Colistin and tigecycline were the most effective antimicrobial agents with 90.3% and 82.4% susceptibility. Notably, 16 (9.7%) isolates showed resistance to colistin. Overall, 33 (20%), 31 (18.8%), and 95 (57.6%) isolates were categorized as strong, moderate, and weak biofilm-producer, respectively. Additionally, blaTEM, blaSHV, blaCTX−M, blaNDM−1, and blaNDM−6 resistance genes were detected in 98 (59.4%), 54 (32.7%), 77 (46.7%), 3 (1.8%), and 3 (1.8%) isolates, respectively. Inactivation of mgrB gene due to nonsense mutations and insertion of IS elements was observed in 6 colistin resistant isolates. Colistin resistance was found to be linked to upregulation of pmrA-K, phoP, and phoQ genes. Three of blaNDM−1 and 3 of blaNDM−6 variants were found to be carried by IncL/M and IncF plasmid, respectively. MLST revealed that blaNDM positive isolates were clonally related and belonged to three distinct clonal complexes, including ST147, ST15 and ST3299. Conclusions The large-scale surveillance and effective infection control measures are also urgently needed to prevent the outbreak of diverse carbapenem- and colistin- resistant isolates in the future.

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Healthcare personnel experiences implementing carbapenem-resistant Enterobacterales infection control measures at a ventilator-capable skilled nursing facility—A qualitative analysis

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2021.265 ◽

2021 ◽

pp. 1-7

Author(s):

Katharina R. Rynkiewich ◽

Jinal Makhija ◽

Mary Carl M. Froilan ◽

Ellen C. Benson ◽

Alice Han ◽

...

Keyword(s):

Infection Control ◽

Multidrug Resistant ◽

Control Measures ◽

Healthcare Personnel ◽

Skilled Nursing ◽

Infection Control Measures ◽

Carbapenem Resistant ◽

Resident Care ◽

Semistructured Interviews ◽

Self Protection

Abstract Objective: Ventilator-capable skilled nursing facilities (vSNFs) are critical to the epidemiology and control of antibiotic-resistant organisms. During an infection prevention intervention to control carbapenem-resistant Enterobacterales (CRE), we conducted a qualitative study to characterize vSNF healthcare personnel beliefs and experiences regarding infection control measures. Design: A qualitative study involving semistructured interviews. Setting: One vSNF in the Chicago, Illinois, metropolitan region. Participants: The study included 17 healthcare personnel representing management, nursing, and nursing assistants. Methods: We used face-to-face, semistructured interviews to measure healthcare personnel experiences with infection control measures at the midpoint of a 2-year quality improvement project. Results: Healthcare personnel characterized their facility as a home-like environment, yet they recognized that it is a setting where germs were ‘invisible’ and potentially ‘threatening.’ Healthcare personnel described elaborate self-protection measures to avoid acquisition or transfer of germs to their own household. Healthcare personnel were motivated to implement infection control measures to protect residents, but many identified structural barriers such as understaffing and time constraints, and some reported persistent preference for soap and water. Conclusions: Healthcare personnel in vSNFs, from management to frontline staff, understood germ theory and the significance of multidrug-resistant organism transmission. However, their ability to implement infection control measures was hampered by resource limitations and mixed beliefs regarding the effectiveness of infection control measures. Self-protection from acquiring multidrug-resistant organisms was a strong motivator for healthcare personnel both outside and inside the workplace, and it could explain variation in adherence to infection control measures such as a higher hand hygiene adherence after resident care than before resident care.

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