Molecular Detection of Carbapenemase-Encoding Genes in Multidrug-Resistant Acinetobacter baumannii Clinical Isolates in South Africa

Introduction. Carbapenem-resistant Acinetobacter baumannii has been responsible for an increasing number of hospital-acquired infections globally. The study investigated the prevalence of carbapenemase-encoding genes in clinical multidrug-resistant A. baumannii strains. Materials and Methods. A total of 100 nonduplicate multidrug-resistant A. baumannii strains were cultured from clinical samples obtained from healthcare facilities in the O. R. Tambo district. The strains were confirmed by detecting the intrinsic blaOXA-51-like gene. Antimicrobial susceptibility testing was performed by VITEK® 2 and autoSCAN-4 systems. The MIC of imipenem and meropenem was rechecked by E-test. Colistin MIC was confirmed by the broth microdilution method. Real-time PCR was performed to investigate the presence of carbapenemase-encoding genes. Results. Most strains showed high resistance rates (>80%) to the antibiotics tested. Resistance to amikacin, tetracycline, and tigecycline were 50%, 64%, and 48%, respectively. All strains were fully susceptible to colistin. The blaOXA-51-like was detected in all strains whilst blaOXA-23-like, blaOXA-58-like, blaOXA-24-like, blaIMP-1, blaVIM, and blaNDM-1 were found in 70%, 8%, 5%, 4%, 3%, and 2% of strains, respectively. None of the tested strains harboured the genes blaSIM and blaAmpC. The coexistence of blaOXA-23-like, and blaIMP-1 or blaOXA-58-like was detected in 1% and 2% strains, respectively. A distinct feature of our findings was the coharbouring of the genes blaOXA-23-like, blaOXA-58-like, and blaIMP-1 in 2% strains, and this is the first report in the Eastern Cape Province, South Africa. The intI1 was carried in 80% of tested strains whilst ISAba1/blaOXA-51-like and ISAba1/blaOXA-23-like were detected in 15% and 40% of the strains, respectively. The detection of blaOXA-23-like, ISAba1/blaOXA-51-like, ISAba1/blaOXA-23-like, and blaOXA-23-like, blaOXA-58-like, and blaIMP-1 carbapenemases in strains had a significant effect on both imipenem and meropenem MICs. Conclusions. Results showed a high level of oxacillinases producing A. baumannii circulating in our study setting, highlighting the need for local molecular surveillance to inform appropriate management and prevention strategies.

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Prevalence of β-lactamase-encoding genes and molecular typing of Acinetobacter baumannii isolates carrying carbapenemase OXA-24 in children

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-021-00480-5 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Neda Yousefi Nojookambari ◽

Mehrzad Sadredinamin ◽

Razieh Dehbanipour ◽

Zohreh Ghalavand ◽

Gita Eslami ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Genetic Relatedness ◽

Medical Center ◽

Resistance Rate ◽

Diffusion Method ◽

Microdilution Method ◽

Broth Microdilution Method ◽

Antimicrobial Susceptibility Tests ◽

Susceptibility Tests ◽

Encoding Genes

Abstract Background β-Lactam antibiotics have been broadly used for the treatment of Acinetobacter baumannii infections, resulting in development of β-lactam inactivating β-lactamases. Here, we described antibiotic resistance rate, prevalence of β-lactamase-encoding genes, and clonal relationships of A. baumannii strains isolated from children referred to Children’s Medical Center in Tehran, Iran, during 2019–2020. Methods A total of 60 non-replicate A. baumannii isolates were recovered from clinical specimens of pediatric patients. Antibiotic susceptibility testing was done by the disc diffusion method. Colistin susceptibility of isolates was performed by the broth microdilution method. β-lactamase-encoding genes were characterized by PCR. The presence of ISAba1 element upstream of the several oxacillinase genes was also checked. Genetic relatedness of isolates was determined by using random amplification of polymorphic DNA (RAPD) typing. Results The antimicrobial susceptibility tests showed that 83.3% of A. baumannii isolates were MDR, and 40% XDR. Both MDR and XDR A. baumannii isolates were susceptible to colistin. The frequency of blaOXA-51-like, blaOXA-23-like, blaTEM, blaOXA-24-like, blaPER, blaSHV, blaCTX-M, blaOXA-58-like, and blaIMP was 100, 93.33, 60, 36.67, 28.33, 8.33, 5, 3.33, and 1.67%, respectively. Coexistence of ISAba1/blaOXA-23-like and ISAba1/blaOXA-51-like was observed in 65% and 85% of isolates, respectively. RAPD analysis revealed 4 common types and 2 single types of A. baumannii isolates. Conclusions The multiple clones harboring blaOXA-23-like, ISAba1-blaOXA-51-like, and ISAba1-blaOXA-23-like were responsible for the spread of A. baumannii isolates in our clinical wards. Dissemination of the well-established clones is worrisome and would become therapeutic challenges due to the possible transferring genetic elements associated with resistance.

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Multidrug Resistant Acinetobacter baumannii Biofilms: Evaluation of Phenotypic–Genotypic Association and Susceptibility to Cinnamic and Gallic Acids

Frontiers in Microbiology ◽

10.3389/fmicb.2021.716627 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mahmoud M. Sherif ◽

Walid F. Elkhatib ◽

Wafaa S. Khalaf ◽

Nooran S. Elleboudy ◽

Neveen A. Abdelaziz

Keyword(s):

Acinetobacter Baumannii ◽

Genetic Basis ◽

Multidrug Resistant ◽

Antimicrobial Activities ◽

Cinnamic Acids ◽

Microdilution Method ◽

Broth Microdilution Method ◽

Life Threatening ◽

Genotypic Association ◽

Polymerase Chain

Acinetobacter baumannii armed with multidrug resistance (MDR) and biofilm-forming ability is increasingly recognized as an alarming pathogen. A deeper comprehension of the correlation between these two armories is required in circumventing its infections. This study examined the biofilm-forming ability of the isolates by crystal violet staining and the antibiotic susceptibility by broth microdilution method. The genetic basis of the MDR and biofilm-forming phenotypes was screened by polymerase chain reaction. The antimicrobial activities of cinnamic and gallic acids against planktonic cells and biofilms of A. baumannii were investigated, and the findings were confirmed with scanning electron microscopy (SEM). Among 90 A. baumannii isolates, 69 (76.6%) were MDR, and all were biofilm formers; they were classified into weak (12.2%), moderate (53.3%), and strong (34.5%) biofilm formers. Our results underlined a significant association between MDR and enhanced biofilm formation. Genotypically, the presence of blaVIM and blaOXA–23 genes along with biofilm-related genes (ompA, bap, and csuE) was statistically associated with the biofilm-forming abilities. Impressively, both gallic and cinnamic acids could significantly reduce the MDR A. baumannii biofilms with variable degrees dependent on the phenotype–genotype characteristics of the tested isolates. The current findings may possess future therapeutic impact through augmenting antimicrobial arsenal against life-threatening infections with MDR A. baumannii biofilms.

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Multidrug-Resistant Listeria Species Shows Abundance in Environmental Waters of a Key District Municipality in South Africa

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18020481 ◽

2021 ◽

Vol 18 (2) ◽

pp. 481

Author(s):

Liyabona Mpondo ◽

Kingsley Ehi Ebomah ◽

Anthony Ifeanyi Okoh

Keyword(s):

South Africa ◽

Surface Waters ◽

Multidrug Resistant ◽

Eastern Cape ◽

Environmental Waters ◽

Potential Health ◽

Phenotypic Resistance ◽

Listeria Species ◽

Polymerase Chain ◽

Identification And Characterization

The prevalence of bacteria with multidrug-resistance (MDR) is a significant threat to public health globally. Listeria spp. are naturally ubiquitous, with L. monocytogenes particularly being ranked as important foodborne disease-causing microorganisms. This study aimed to evaluate the incidence and determine the antimicrobial resistance (AMR) profiles of multidrug-resistant Listeria spp. (MDRL) isolated from different environmental samples (river and irrigation water) in the Sarah Baartman District Municipality (SBDM), Eastern Cape Province (ECP), South Africa. Molecular identification and characterization were carried out using polymerase chain reaction (PCR) and isolates that exhibited phenotypic resistance were further screened for relevant antimicrobial-resistant genes (ARGs). Findings revealed a total of 124 presumptive Listeria isolates; 69 were molecularly confirmed Listeria species. Out of the confirmed species, 41 isolates (59%) were classified as L. monocytogenes while 9 (13%) were classified as L. welshimeri. All Listeria spp. exhibited phenotypic resistance against ampicillin, penicillin, and trimethoprim-sulphamethoxazole and further screening revealed ARGs in the following proportions: sulI (71%), blaTEM (66%), tetA (63%), and blaCIT (33%). Results confirmed the occurrence of ARGs among Listeria inhabiting surface waters of ECP. The present study indicates that the river water samples collected from SBDM are highly contaminated with MDRL, hence, constituting a potential health risk.

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1579. Multidrug-Resistant Candida auris Isolates From New York Hospitals and Healthcare Facilities Are Susceptible to Antifungal Combinations

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.1443 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S576-S577

Author(s):

Brittany O’Brien ◽

Sudha Chaturvedi ◽

Vishnu Chaturvedi

Keyword(s):

New York ◽

Diagnostic System ◽

Multidrug Resistant ◽

Drug Combinations ◽

Antifungal Drugs ◽

Drug Resistant ◽

Candida Auris ◽

Current Case ◽

Healthcare Facilities ◽

Broth Microdilution Method

Abstract Background Candida auris outbreak continues unabated in New York with the current case counts exceeding 300 patients. We used a modification of standard CLSI broth microdilution method (BMD) if two-drug combinations are efficacious against C. auris isolates with high-resistance to fluconazole (FZ, MIC50 >256 mg/L), and variable resistance to other broad-spectrum antifungal drugs. Methods BMD plates were custom-designed and quality controlled by TREK Diagnostic System. The combination tests of 15 drug-resistant C. auris involved microtiter wells with the initial 144 two-drug combinations and their two-fold dilutions (1/2–1/32) to get 864 two-drug combinations finally. We utilized MIC100 endpoints for the drug combination readings as reported earlier for the intra- and inter-laboratory agreements obtained against Candida species and Aspergillus fumigatus (Antimicrob Agents Chemother. 2015. 59:1759–1766). We also tested minimum fungicidal concentrations (MFC). Results We tested all possible 864 two-drug antifungal combinations for nine antifungal drugs in use to yield 12,960 MIC100 readings, and MFC readings for 15 C. auris isolates. Flucytosine (FLC) at 2.0 mg/L potentiated most successful combinations with other drugs. Micafungin (MFG), Anidulafungin (AFG), Caspofungin (CAS) at individual concentrations of 0.25 mg/L combined well with FLC (2.0 mg/L) to yield MIC100 for 14, 13, and 12 of 15 C. auris isolates tested, respectively. MFG/FLC combination was also fungicidal for 4 of 15 isolates. AMB / FLC (0.25/1.0 mg/L) yielded MIC100 for 13 isolates and MFC for three test isolates. Posaconazole (POS), and Isavuconazole (ISA) and Voriconazole (VRC) also combined well with FLC (0.25/2.0 mg/L) to yield MIC100 for 12, 13, and 13 isolates, respectively. POS/FLC combination was fungicidal for three isolates. Conclusion We identified seven two drug-combinations of antifungals efficacious against drug-resistant C. auris strains. The modified BMD combination susceptibility testing could be used by the clinical laboratories to assist providers with the selection of optimal treatment for C. auris candidemia. Disclosures All authors: No reported disclosures.

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Trends in Antifungal Drug Susceptibility of Cryptococcus neoformans Isolates Obtained through Population-Based Surveillance in South Africa in 2002-2003 and 2007-2008

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00048-11 ◽

2011 ◽

Vol 55 (6) ◽

pp. 2606-2611 ◽

Cited By ~ 43

Author(s):

Nelesh P. Govender ◽

Jaymati Patel ◽

Marelize van Wyk ◽

Tom M. Chiller ◽

Shawn R. Lockhart ◽

...

Keyword(s):

South Africa ◽

Cryptococcus Neoformans ◽

Amphotericin B ◽

Fungal Infections ◽

Antifungal Susceptibility ◽

Drug Susceptibility ◽

Population Based ◽

Content Type ◽

Microdilution Method ◽

Broth Microdilution Method

ABSTRACTCryptococcus neoformansis the most common cause of meningitis among adult South Africans with HIV infection/AIDS. Widespread use of fluconazole for treatment of cryptococcal meningitis and other HIV-associated opportunistic fungal infections in South Africa may lead to the emergence of isolates with reduced fluconazole susceptibility. MIC testing using a reference broth microdilution method was used to determine if isolates with reduced susceptibility to fluconazole or amphotericin B had emerged among cases of incident disease. Incident isolates were tested from two surveillance periods (2002-2003 and 2007-2008) when population-based surveillance was conducted in Gauteng Province, South Africa. These isolates were also tested for susceptibility to flucytosine, itraconazole, voriconazole, and posaconazole. Serially collected isolate pairs from cases at several large South African hospitals were also tested for susceptibility to fluconazole. Of the 487 incident isolates tested, only 3 (0.6%) demonstrated a fluconazole MIC of ≥16 μg/ml; all of these isolates were from 2002-2003. All incident isolates were inhibited by very low concentrations of amphotericin B and exhibited very low MICs to voriconazole and posaconazole. Of 67 cases with serially collected isolate pairs, only 1 case was detected where the isolate collected more than 30 days later had a fluconazole MIC value significantly higher than the MIC of the corresponding incident isolate. Although routine antifungal susceptibility testing of incident isolates is not currently recommended in clinical settings, it is still clearly important for public health to periodically monitor for the emergence of resistance.

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Perceptions of nursing students regarding responsible use of social media in the Eastern Cape

Curationis ◽

10.4102/curationis.v38i2.1496 ◽

2015 ◽

Vol 38 (2) ◽

Cited By ~ 21

Author(s):

Thando Nyangeni ◽

Suzette Du Rand ◽

Dalena Van Rooyen

Keyword(s):

South Africa ◽

Social Media ◽

Nursing Education ◽

Nursing Students ◽

Research Study ◽

Structured Interview ◽

Eastern Cape ◽

Clinical Environment ◽

Healthcare Facilities ◽

Use Of Social Media

Background: Social media have become a popular communication system that has transformed communication from the traditional to the Web-based model. Because social media use has no limitations to place and time, it is now used extensively at clinical facilities. Social media useis becoming a popular activity amongst students at Nursing Education Institutions (NEI) in South Africa. However, lack of accountability and unethical use of social media by nursing students in South Africa has been reported.Objectives: The aim of the study was to explore and describe the perceptions of nursing students regarding responsible use of social media.Methods: A qualitative, descriptive, explorative and contextual research design was used to explore and describe the perceptions of nursing students regarding the responsible use of social media. Twelve nursing students registered for the undergraduate nursing degree were purposely selected and interviewed individually using a semi-structured interview method.Results: The results of this research study demonstrate that nursing students use socialmedia irresponsibly. Nursing students experience blurred boundaries between personal and professional lines and lack accountability when using social media.Conclusion: The extensive use of social media in the clinical environment, by healthcare students, requires a joint effort by Nursing Education Institutions and healthcare facilities to ensure that social media are used in an ethically acceptable manner. The implementation of the recommendations of this research study could positively influence legally and ethically acceptable use of social media at healthcare facilities.

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528. Longitudinal Evaluation of Chlorhexidine Resistance Among Multidrug-Resistant Organisms in Relation to Intervention of Daily Chlorhexidine Bathing in Adult Intensive Care Units

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.597 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S254-S254

Author(s):

Min Ja Kim ◽

You Seung Chung ◽

Hojin Lee ◽

Jin Woong Suh ◽

Yoojung Cheong ◽

...

Keyword(s):

Intensive Care ◽

Intensive Care Units ◽

Clinical Laboratory ◽

Multidrug Resistant ◽

University Hospital ◽

Microdilution Method ◽

Broth Microdilution Method ◽

Multidrug Resistant Organisms ◽

Before And After ◽

The Republic

Abstract Background Chlorhexidine digluconate (CHG), the most widely used antiseptic, has recently been applied to patient washing to decolonize the multidrug-resistant organisms (MDROs), but there are little data on susceptibilities of MDROs to CHG. The purpose of this study was to evaluate CHG resistance among MDROs before and after the intervention of daily CHG bathing in adult intensive care units (ICUs). Methods The intervention of daily body washing with 2% CHG cloths were taken in adult patients the medical or surgical ICU of 23-bed by a crossover manner for 6 months (MICU, July to December 2017; SICU, January to June 2018) in a 1,050-bed, university hospital in the Republic of Korea. Available MDRO isolates were randomly selected from clinical cultures of ICU patients within 6 months before, during and after the intervention, including MRSA, MR-CoNS, VRE, Carbapenem-resistant Pseudomonas aeruginosa (CR-PA), CR-Acinetobacter baumannii (CR-AB). Minimum inhibitory concentrations (MICs) were determined using the broth microdilution method set by the Clinical Laboratory Standards Institute. Determination of the minimum bactericidal concentrations (MBCs) was performed by subculturing 10 µL from each well without visible microbial growth. Cumulative amounts of CHG used in both ICUs was estimated across the study period from January 2008 to June 2018. Results The cumulative CHG consumption from both ICUs increased sharply from 27,503 g to 29,556 g after one-year intervention. The ranges of MICs and MBCs of CHG among MDRO clinical isolates selected by a 6-month phase are summarized in Table 1. Particularly, CR-PA and CR-AB isolates revealed four to eight times higher MICs and MBCs compared with the majority of Gram-positives excepting some VRE isolates. On the other hand, neither MICs and MBCs ranges of CHG from the MDRO isolates nor the monthly incidence of the MDROs from both ICUs were significantly increased before and after the intervention of daily CHG bathing. Conclusion This study indicates that some Gram-negative MDRO isolates with higher MICs and MBCs of CHG might be from longstanding exposure to CHG or efflux pumps. Although 2% daily CHG bathing uses over 1,000 times higher concentrations than the lethal concentration, it might be needed to monitor CHG resistance among MDROs. Disclosures All authors: No reported disclosures.

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Curcumin, a Natural Antimicrobial Agent with Strain-Specific Activity

Pharmaceuticals ◽

10.3390/ph13070153 ◽

2020 ◽

Vol 13 (7) ◽

pp. 153

Author(s):

Artur Adamczak ◽

Marcin Ożarowski ◽

Tomasz M. Karpiński

Keyword(s):

Inhibitory Concentration ◽

Specific Activity ◽

Curcuma Longa ◽

Antimicrobial Properties ◽

Antiviral Agent ◽

Multidrug Resistant ◽

Gram Negative Bacteria ◽

Microdilution Method ◽

Broth Microdilution Method ◽

Bioactive Substance

Curcumin, a principal bioactive substance of turmeric (Curcuma longa L.), is reported as a strong antioxidant, anti-inflammatory, antibacterial, antifungal, and antiviral agent. However, its antimicrobial properties require further detailed investigations into clinical and multidrug-resistant (MDR) isolates. In this work, we tested curcumin’s efficacy against over 100 strains of pathogens belonging to 19 species. This activity was determined by the broth microdilution method and by calculating the minimum inhibitory concentration (MIC). Our findings confirmed a much greater sensitivity of Gram-positive than Gram-negative bacteria. This study exhibited a significantly larger variation in the curcumin activity than previous works and suggested that numerous clinical strains of widespread pathogens have a poor sensitivity to curcumin. Similarly, the MICs of the MDR types of Staphylococcus aureus, S. haemolyticus, Escherichia coli, and Proteus mirabilis were high (≥2000 µg/mL). However, curcumin was effective against some species and strains: Streptococcus pyogenes (median MIC = 31.25 µg/mL), methicillin-sensitive S. aureus (250 µg/mL), Acinetobacter lwoffii (250 µg/mL), and individual strains of Enterococcus faecalis and Pseudomonas aeruginosa (62.5 µg/mL). The sensitivity of species was not associated with its affiliation to the genus, and it could differ a lot (e.g., S. pyogenes, S. agalactiae and A. lwoffii, A. baumannii). Hence, curcumin can be considered as a promising antibacterial agent, but with a very selective activity.

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In vitro activity of the trinem sanfetrinem (GV104326) against gram-positive organisms.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.9.2142 ◽

1996 ◽

Vol 40 (9) ◽

pp. 2142-2146 ◽

Cited By ~ 11

Author(s):

K V Singh ◽

T M Coque ◽

B E Murray

Keyword(s):

Staphylococcus Aureus ◽

Vitro Activity ◽

In Vitro Activity ◽

Gram Positive ◽

Gram Positive Bacteria ◽

Microdilution Method ◽

Broth Microdilution Method ◽

Gentamicin Resistance ◽

High Level

The in vitro activity of the trinem sanfetrinem (formerly GV104326) (GV) was compared with that of vancomycin, ampicillin, and/or nafcillin against 287 gram-positive bacteria, including methicillin-resistant Staphylococcus aureus and multiresistant enterococci, by the agar and microbroth dilution methods. GV demonstrated 2 to 16 times more activity than ampicillin and nafcillin against the majority of these organisms. The MIC range of GV was 16 to 64 micrograms/ml for 19 Enterococcus faecium strains that were highly resistant to ampicillin (ampicillin MIC range, 64 to 512 micrograms/ml) and vancomycin resistant and 0.25 to 32 micrograms/ml for resistant Rhodococcus spp. Similar activities (+/-1 dilution) were observed by either the agar or the broth microdilution method. GV demonstrated bactericidal activity against a beta-lactamase-producing Enterococcus faecalis strain and against two methicillin-susceptible Staphylococcus aureus strains in 10(5)-CFU/ml inocula. Synergy between GV and gentamicin was observed against an E. faecalis strain that lacked high-level gentamicin resistance. The activity of GV suggests this compound warrants further study.

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Genetic Basis of Multidrug Resistance in Acinetobacter baumannii Clinical Isolates at a Tertiary Medical Center in Pennsylvania

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00570-08 ◽

2008 ◽

Vol 52 (11) ◽

pp. 3837-3843 ◽

Cited By ~ 95

Author(s):

Jennifer M. Adams-Haduch ◽

David L. Paterson ◽

Hanna E. Sidjabat ◽

Anthony W. Pasculle ◽

Brian A. Potoski ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Genetic Basis ◽

Medical Center ◽

Multidrug Resistant ◽

Quinolone Resistance ◽

Clinical Isolates ◽

Clinical Microbiology Laboratory ◽

Carbapenemase Gene ◽

Methylase Gene ◽

High Level

ABSTRACT A total of 49 unique clinical isolates of multidrug-resistant (MDR) Acinetobacter baumannii identified at a tertiary medical center in Pittsburgh, Pennsylvania, between August 2006 and September 2007 were studied for the genetic basis of their MDR phenotype. Approximately half of all A. baumannii clinical isolates identified during this period qualified as MDR, defined by nonsusceptibility to three or more of the antimicrobials routinely tested in the clinical microbiology laboratory. Among the MDR isolates, 18.4% were resistant to imipenem. The frequencies of resistance to amikacin and ciprofloxacin were high at 36.7% and 95.9%, respectively. None of the isolates was resistant to colistin or tigecycline. The presence of the carbapenemase gene bla OXA-23 and the 16S rRNA methylase gene armA predicted high-level resistance to imipenem and amikacin, respectively. bla OXA-23 was preceded by insertion sequence ISAba1, which likely provided a potent promoter activity for the expression of the carbapenemase gene. The structure of the transposon defined by ISAba1 differed from those reported in Europe, suggesting that ISAba1-mediated acquisition of bla OXA-23 may occur as an independent event. Typical substitutions in the quinolone resistance-determining regions of the gyrA and parC genes were observed in the ciprofloxacin-resistant isolates. Plasmid-mediated quinolone resistance genes, including the qnr genes, were not identified. Fifty-nine percent of the MDR isolates belonged to a single clonal group over the course of the study period, as demonstrated by pulsed-field gel electrophoresis.

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