Molecular Characterizations, Virulence Determinants and Antimicrobial Resistance Profiles of Methicillin-Resistant Staphylococcus aureus (MRSA) in the North of Iran

Abstract Background: Emergence and prevalence of Methicillin-Resistant Staphylococcus aureus (MRSA) has become a major universal health concern, limiting therapeutic options. Methods: In the North side of Iran, during the years 2015 to 2017, a total number of 37 MRSA isolates, including 19 clinical isolates from hospitalized patients and 18 colonizing isolates from health care workers were identified from three hospitals, in Gorgan, North of Iran. Antimicrobial susceptibility test was performed using the disk diffusion method and E-test. The presence of virulence and antibiotic resistance determinants were evaluated by PCR. The genotypic characterization was further analyzed using multi-locus sequence, spa, SCCmec, and agr typing.Results: The frequency of MRSA among S. aureus isolates was 38.14% (37/97). The most frequent S. aureus resistant isolates were found to be obstinate against penicillin (98%) and gentamicin (82.5%). Additionally, the lowest resistance rates were found against daptomycin (0%), vancomycin (2.7%), and quinupristin-dalfopristin (5.4%). All MRSA isolates were susceptible to daptomycin with MIC50/MIC90 of 0.25/0.5 µg/ml. One isolate belonging to the ST239-SCCmecIII/t037 clone (MIC≥16μg/ml) was resistant to vancomycin. All but one isolate that shares the ST22-SCCmec IV/t790 strain were positive for both tsst and pvl genes. The most predominant MRSA isolates (27%) were associated with the ST239-SCCmec III/t037 clone; and followed by ST239-SCCmec III/t924 (16.2%).Conclusions: In our study, circulating MRSA strains were genetically diverse with a high prevalence of the ST239-SCCmecIII/t037 clone. These findings emphasize the need for future and continuous surveillance studies on MRSA to prevent the dissemination of multidrug resistance and existing MRSA clones in an effective manner.

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Molecular Characterization, Virulence Determinants, and Antimicrobial Resistance Profile of Methicillin-Resistant Staphylococcus aureus in the North of Iran; a High Prevalence of ST239-SCCmec III/t037 Clone

Chemotherapy ◽

10.1159/000520482 ◽

2021 ◽

pp. 1-10

Author(s):

Hao Ying ◽

Trias Mahmudiono ◽

Tawfeeq Alghazali ◽

Walid Kamal Abdelbasset ◽

Parand Khadivar ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Health Concern ◽

Diffusion Method ◽

Virulence Determinants ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

The North ◽

Effective Manner ◽

High Prevalence ◽

North Of Iran

Objectives: Emergence and prevalence of methicillin-resistant Staphylococcus aureus (MRSA) have become a major universal health concern, limiting therapeutic options. Methods: A total number of 37 MRSA isolates, including 19 clinical isolates from hospitalized patients and 18 colonizing isolates from health care workers were identified from 3 hospitals, in Gorgan, North of Iran. Antimicrobial susceptibility test was performed using the disk diffusion method and E-test. The presence of virulence and antibiotic resistance determinants were evaluated by PCR. The genotypical characterization was further analyzed using multi-locus sequence, spa, staphylococcal cassette chromosome, mec (SCCmec), and agr typing. Results: The frequency of MRSA among S. aureus isolates was 38.14% (37/97). The most frequent S. aureus resistant isolates were found to be obstinate against penicillin (98%) and gentamicin (82.5%). Additionally, the lowest resistance rates were found against daptomycin (0%), vancomycin (2.7%), and quinupristin-dalfopristin (5.4%). All MRSA isolates were susceptible to daptomycin with minimum inhibitory concentration (MIC)50/MIC90 of 0.25/0.5 μg/mL. One isolate belonging to sequence type 239 (ST239)-SCCmecIII/t037 clone (MIC ≥16 μg/mL) was resistant to vancomycin. All but 1 isolate that shares ST22-SCCmec IV/t790 strain were positive for both tsst and pvl genes. The most predominant MRSA isolates (27%) were associated with ST239-SCCmec III/t037, and ST239-SCCmec III/t924 (16.2%) clones, subsequently. In our study, circulating MRSA strains were genetically diverse with a high prevalence of ST239-SCCmec III/t037 clone. Conclusion: These findings emphasize the need for future and continuous surveillance studies on MRSA to prevent the dissemination of existing multidrug resistance MRSA clones in an effective manner.

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Livestock-Associated Methicillin-Resistant Staphylococcus aureus (MRSA) in Purulent Subcutaneous Lesions of Farm Rabbits

Foods ◽

10.3390/foods9040439 ◽

2020 ◽

Vol 9 (4) ◽

pp. 439 ◽

Cited By ~ 1

Author(s):

Vanessa Silva ◽

Telma de Sousa ◽

Paula Gómez ◽

Carolina Sabença ◽

Madalena Vieira-Pinto ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agents ◽

Methicillin Resistant Staphylococcus Aureus ◽

Multidrug Resistant ◽

Diffusion Method ◽

Spa Typing ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Resistance To Penicillin ◽

Mrsa Strains

Methicillin-resistant Staphylococcus aureus (MRSA) are one of the main pathogens associated with purulent infections. MRSA clonal complex 97 (CC97) has been identified in a wide diversity of livestock animals. Therefore, we aimed to investigate the antibiotic resistance profiles of MRSA strains isolated from purulent lesions of food-producing rabbits. Samples from purulent lesions of 66 rabbits were collected in a slaughterhouse in Portugal. Samples were seeded onto ORSAB plates with 2 mg/L of oxacillin for MRSA isolation. Susceptibility to antibiotics was tested by the disk diffusion method against 14 antimicrobial agents. The presence of resistance genes, virulence factors and the immune evasion cluster (IEC) system was studied by polymerase chain reaction. All isolates were characterized by multilocus sequence typing (MLST), agr and spa typing. From the 66 samples analyzed, 16 (24.2%) MRSA were detected. All strains were classified as multidrug-resistant as they were resistant to at least three classes of antibiotics. All isolates showed resistance to penicillin, erythromycin and clindamycin. Seven isolates were resistant to gentamicin and harbored the aac(6′)-Ie-aph (2″)-Ia gene. Resistance to tetracycline was detected in 10 isolates harboring the tet(K) gene. The IEC genes were detected in three isolates. MRSA strains belonged to CC97, CC1, CC5, CC15 or CC22. The isolates were assigned to six different spa types. In this study we found a moderate prevalence of multidrug-resistant MRSA strains in food-producing rabbits. This may represent concern for food safety and public health, since cross-contamination may occur, leading to the spread of MRSA and, eventually, the possibility of ingestion of contaminated meat.

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Outbreak of Methicillin-Resistant Staphylococcus aureus with Reduced Susceptibility to Glycopeptides in a Parisian Hospital

Journal of Clinical Microbiology ◽

10.1128/jcm.38.8.2985-2988.2000 ◽

2000 ◽

Vol 38 (8) ◽

pp. 2985-2988 ◽

Cited By ~ 58

Author(s):

François Guerin ◽

Annie Buu-Hoï ◽

Jean-Luc Mainardi ◽

Guillaume Kac ◽

Nathalie Colardelle ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Brain Heart Infusion ◽

Multidrug Resistant ◽

Pulsed Field Gel Electrophoresis ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Mrsa Strains ◽

Standard Disk

Epidemiological relationships were investigated between 40 methicillin-resistant Staphylococcus aureus (MRSA) strains with decreased glycopeptide susceptibility isolated from November 1998 to March 1999 from 39 patients (17 infected and 22 colonized patients) in nine wards of the Broussais Hospital, Paris, France. Reduced glycopeptide susceptibility was readily detected on brain heart infusion (BHI) agar containing 6 μg of teicoplanin per ml and on gradient plates, but not by the standard disk diffusion method. The MICs of vancomycin and teicoplanin, determined on BHI agar, were 4 and 8 to 32 μg/ml, respectively (standard antibiotic dilution), and 4 to 8 and 8 to 32 μg/ml, respectively (E-test). All strains were resistant to macrolides, aminoglycosides, tetracycline, rifampin, sulfonamides, and pefloxacin, showed reduced susceptibility to fusidic acid and fosfomycin, and were susceptible to trimethoprim and chloramphenicol. Pulsed-field gel electrophoresis and lysotyping revealed that a multidrug-resistant MRSA clone with decreased susceptibility to glycopeptides has been discretely endemic since at least 1996 in our institution, where it was responsible for an outbreak in November and December 1998.

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Prevalence and molecular epidemiology of ceftaroline non-susceptible methicillin-resistant Staphylococcus aureus isolates, first clinical report from Iran

Acta Microbiologica et Immunologica Hungarica ◽

10.1556/030.2020.01273 ◽

2020 ◽

Author(s):

Amin Khoshbayan ◽

Aref Shariati ◽

Ehsanollah Ghaznavi-Rad ◽

Alex van Belkum ◽

Davood Darban-Sarokhalil

Keyword(s):

Staphylococcus Aureus ◽

Tracheal Aspirate ◽

Generation Cephalosporin ◽

Diffusion Method ◽

Type I ◽

Clinical Report ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Novel Approach ◽

Mrsa Strains

AbstractBackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens in Iran with a high prevalence and a high level of antibiotic resistance. Ceftaroline is a fifth generation cephalosporin binding and inhibiting penicillin binding protein (PBP2a).MethodsIn the present study, 228 clinical MRSA isolates were collected from four cities of Iran and their susceptibility to ceftaroline was evaluated by E-test and the disk diffusion method.ResultsOur results showed a high susceptibility rate (97.3%) to ceftaroline in MRSA strains from Iran. Six isolates were found to be ceftaroline non-susceptible (CPT-NS) with Minimum inhibitory concentration (MIC) ≥2 µg/mL. All CPT-NS isolates were isolated from blood and tracheal aspirate and belonged to SCCmec type III as well as agr type I and were all susceptible to vancomycin. Out of six isolates, three, two and one belonged to spa type t030, t4864, and t969, respectively. Vancomycin, quinupristin/dalfopristin, linezolid, chloramphenicol, and tigecycline were the most active agents against CPT-NS isolates.ConclusionDue to the broad-spectrum activity and low toxicity of ceftaroline as well as the increased rate of vancomycin resistance among MRSA strains in recent years, ceftaroline can be considered as a novel approach to treat MRSA-induced infections.

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Predominance of PVL-negative community-associated methicillin-resistant Staphylococcus aureus sequence type 8 in newly diagnosed HIV-infected adults, Tanzania

European Journal of Clinical Microbiology & Infectious Diseases ◽

10.1007/s10096-021-04160-2 ◽

2021 ◽

Author(s):

Joel Manyahi ◽

Sabrina J. Moyo ◽

Said Aboud ◽

Nina Langeland ◽

Bjørn Blomberg

Keyword(s):

Staphylococcus Aureus ◽

Hiv Infection ◽

Diffusion Method ◽

Molecular Characteristics ◽

People Living With Hiv ◽

Newly Diagnosed ◽

Beta Lactam ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Inducible Clindamycin Resistance

AbstractDifficult-to-treat infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are of concern in people living with HIV infection as they are more vulnerable to infection. We aimed to identify molecular characteristics of MRSA colonizing newly diagnosed HIV-infected adults in Tanzania. Individuals newly diagnosed with HIV infection were recruited in Dar es Salaam, Tanzania, from April 2017 to May 2018, as part of the randomized clinical trial CoTrimResist (ClinicalTrials.gov identifier: NCT03087890). Nasal/nasopharyngeal isolates of Staphylococcus aureus were susceptibility tested by disk diffusion method, and cefoxitin-resistant isolates were characterized by short-reads whole genome sequencing. Four percent (22/537) of patients carried MRSA in the nose/nasopharynx. MRSA isolates were frequently resistant towards gentamicin (95%), ciprofloxacin (91%), and erythromycin (82%) but less often towards trimethoprim-sulfamethoxazole (9%). Seventy-three percent had inducible clindamycin resistance. Erythromycin-resistant isolates harbored ermC (15/18) and LmrS (3/18) resistance genes. Ciprofloxacin resistance was mediated by mutations of the quinolone resistance-determining region (QRDR) sequence in the gyrA (S84L) and parC (S80Y) genes. All isolates belonged to the CC8 and ST8-SCCmecIV MRSA clone. Ninety-five percent of the MRSA isolates were spa-type t1476, and one exhibited spa-type t064. All isolates were negative for Panton-Valentine leucocidin (PVL) and arginine catabolic mobile element (ACME) type 1. All ST8-SCCmecIV-spa-t1476 MRSA clones from Tanzania were unrelated to the globally successful USA300 clone. Carriage of ST8 MRSA (non-USA300) was common among newly diagnosed HIV-infected adults in Tanzania. Frequent co-resistance to non-beta lactam antibiotics limits therapeutic options when infection occurs.

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Methicillin-resistant Staphylococcus aureus bloodstream infection: risk factors and clinical outcome in non-intensive-care units

Revista da Sociedade Brasileira de Medicina Tropical ◽

10.1590/s0037-86822012000200010 ◽

2012 ◽

Vol 45 (2) ◽

pp. 189-193 ◽

Cited By ~ 4

Author(s):

Karinne Spirandelli Carvalho Naves ◽

Natália Vaz da Trindade ◽

Paulo Pinto Gontijo Filho

Keyword(s):

Risk Factors ◽

Staphylococcus Aureus ◽

Bloodstream Infection ◽

Antimicrobial Agents ◽

Methicillin Resistant Staphylococcus Aureus ◽

Methicillin Resistance ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Staphylococcus Aureus Bloodstream Infection

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is spread out in hospitals across different regions of the world and is regarded as the major agent of nosocomial infections, causing infections such as skin and soft tissue pneumonia and sepsis. The aim of this study was to identify risk factors for methicillin-resistance in Staphylococcus aureus bloodstream infection (BSI) and the predictive factors for death. METHODS: A retrospective cohort of fifty-one patients presenting bacteraemia due to S. aureus between September 2006 and September 2008 was analysed. Staphylococcu aureus samples were obtained from blood cultures performed by clinical hospital microbiology laboratory from the Uberlândia Federal University. Methicillinresistance was determined by growth on oxacillin screen agar and antimicrobial susceptibility by means of the disk diffusion method. RESULTS: We found similar numbers of MRSA (56.8%) and methicillin-susceptible Staphylococcus aureus (MSSA) (43.2%) infections, and the overall hospital mortality ratio was 47%, predominantly in MRSA group (70.8% vs. 29.2%) (p=0.05). Age (p=0.02) was significantly higher in MRSA patients as also was the use of central venous catheter (p=0.02). The use of two or more antimicrobial agents (p=0.03) and the length of hospital stay prior to bacteraemia superior to seven days (p=0.006) were associated with mortality. High odds ratio value was observed in cardiopathy as comorbidity. CONCLUSIONS: Despite several risk factors associated with MRSA and MSSA infection, the use of two or more antimicrobial agents was the unique independent variable associated with mortality.

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Prevalence of Methicillin Resistant and Virulence Determinants in Clinical Isolates of Staphylococcus aureus

The Open Infectious Diseases Journal ◽

10.2174/1874279301810010108 ◽

2018 ◽

Vol 10 (1) ◽

pp. 108-115

Author(s):

Manjunath Chavadi ◽

Rahul Narasanna ◽

Ashajyothi Chavan ◽

Ajay Kumar Oli ◽

Chandrakanth Kelmani. R

Keyword(s):

Staphylococcus Aureus ◽

Cell Morphology ◽

Methicillin Resistance ◽

Protein A ◽

Clinical Isolates ◽

Clinical Samples ◽

Virulence Determinants ◽

Methicillin Resistant ◽

Alternative Medicines ◽

Mrsa Strains

Introduction:Methicillin-resistantStaphylococcus aureus(MRSA) is the major threat that is a result of the uncontrolled use of antibiotics causing a huge loss in health, so understanding their prevalence is necessary as a public health measure.Objective:The aim of this study was to determine the prevalence of methicillin-resistant MRSA and virulence determinant among associatedS. aureusfrom the clinical samples obtained from various hospital and health care centers of the Gulbarga region in India.Materials and Methods:All the collected samples were subjected for the screening ofS. aureusand were further characterized by conventional and molecular methods including their antibiotic profiling. Further, the response of methicillin antibiotic on cell morphology was studied using scanning electron microscopy.Results:A total 126S. aureuswas isolated from the clinical samples which showed, 100% resistant to penicillin, 55.5% to oxacillin, 75.3% to ampicillin, 70.6% to streptomycin, 66.6% to gentamicin, 8.7% to vancomycin and 6.3% to teicoplanin. The selected MRSA strains were found to possessmecA(gene coding for penicillin-binding protein 2A) andfemA(factor essential for methicillin resistance)genetic determinants in their genome with virulence determinants such as Coagulase (coa) and the X region of the protein A (spa)gene. Further, the methicillin response in resistantS. aureusshowed to be enlarged and malformed on cell morphology.Conclusion:The molecular typing of clinical isolates ofS. aureusin this study was highly virulent and also resistant to methicillin; this will assist health professionals to control, exploration of alternative medicines and new approaches to combat Staphylococcal infections more efficiently by using targeted therapy.

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Evaluation of antibiotic resistance pattern and mecA gene frequency in methicillin-resistant Staphylococcus aureus strains collected from clinical specimens in Marand hospital and treatment centers, Iran

Tabari Biomedical Student Research Journal ◽

10.18502/tbsrj.v2i4.5469 ◽

2021 ◽

Author(s):

Abolfazl Jafari-Sales ◽

Zahra Sadeghi Deylamdeh ◽

Afsoon Shariat

Keyword(s):

Staphylococcus Aureus ◽

Antibiotic Resistance ◽

Methicillin Resistant Staphylococcus Aureus ◽

Meca Gene ◽

Disk Diffusion ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Medical Centers ◽

Wide Range

Introduction: Staphylococcus aureus causes a wide range of infections and as a multivalent pathogen is one of the causative agents of nosocomial and community infections. Therefore, the aim of this study was to identify and determine the pattern of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in hospitals and medical centers in Marand city and also to evaluate the presence of mecA gene. Materials and Methods: In this cross-sectional descriptive study, 385 samples of S. aureus were collected from different clinical samples of patients in hospitals and medical centers of Marand city. S. aureus was identified using standard biochemical methods. Methicillin resistance was determined by disk diffusion method in the presence of oxacillin and cefoxitin. The pattern of antibiotic resistance of the strains was determined by disk diffusion method and according to CLSI recommendation and also PCR method was used to evaluate the frequency of MecA gene. Results: In the present study, out of 385 samples of S. aureus, 215 (55.84%) samples were methicillin resistant. PCR results for mecA gene showed that 110 samples had mecA gene. The highest antibiotic resistance was observed against penicillin (100%) and erythromycin (83.63%). Most MRSA were isolated from urine and wound samples. Conclusion: The results of this study indicate the prevalence of methicillin-resistant species and also the increase in antibiotic resistance of MRSA to various antibiotics. Therefore, in order to prevent increased resistance to other antibiotics, it is recommended to avoid inappropriate use of antibiotics.

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Assessment of Mupirocin Resistance among Clinical Isolates of Methicillin Resistant Staphylococcus aureus

10.51429/ejmm30114 ◽

2021 ◽

Vol 30 (1) ◽

pp. 109-114

Author(s):

Nancy M. Attia ◽

Abeer Abd El Rahim Ghazal ◽

Omnia M. Khaleel ◽

Ahmed Gaballah

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Methicillin Resistant ◽

Mupirocin Resistance ◽

High Level ◽

Research Institute Hospital ◽

Low Prevalence ◽

Mrsa Colonization

Background: Methicillin-resistant Staphylococcus aureus (MRSA) colonization is considered a major risk factor for nosocomial infections and its decolonization has reduced these infections. Mupirocin (MUP) is the topical antibiotic of choice for decolonization. MUP decolonization failure is attributed to MUP resistance. Objective: The aim of the current study is to assess MUP resistance among MRSA isolates phenotypically and genotypically. Methodology: Fifty MRSA isolates were identified in Microbiology Department in the Medical Research Institute hospital, Alexandria University. Antibiotic susceptibility to different classes of antibiotics by disk diffusion method was done. MUP minimum inhibitory concentration (MIC) was determined phenotypically by MUP Ezy MIC™ Strips. MUP resistance was determined genetically by multiplex PCR detection of mupA and mupB. Results: Of all MRSA isolates, 6% exhibited high level and none showed low level MUP resistance. Only mupA was detected in all resistant isolates. Conclusion: Despite low prevalence of MUP resistance, it is appropriate to test MUP resistance prior nasal decolonization

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Differentiation of Community-Associated and Livestock-Associated Methicillin-Resistant Staphylococcus aureus Isolates and Identification of spa Types by Use of PCR and High-Resolution Melt Curve Analysis

Journal of Clinical Microbiology ◽

10.1128/jcm.02088-19 ◽

2020 ◽

Vol 58 (5) ◽

Cited By ~ 1

Author(s):

Seyed A. Ghorashi ◽

Jane Heller ◽

Quincy Zhang ◽

Shafi Sahibzada

Keyword(s):

Staphylococcus Aureus ◽

High Resolution ◽

Curve Analysis ◽

Health Concern ◽

Staphylococcal Protein A ◽

Visual Interpretation ◽

High Resolution Melt ◽

Methicillin Resistant ◽

Content Type ◽

Mrsa Strains

ABSTRACT Infections due to methicillin-resistant Staphylococcus aureus (MRSA) are present worldwide and represent a major public health concern. The capability of PCR followed by high-resolution melt (HRM) curve analysis for the detection of community-associated and livestock-associated MRSA strains and the identification of staphylococcal protein A (spa) locus was evaluated in 74 MRSA samples which were isolated from the environment, humans, and pigs on a single piggery. PCR-HRM curve analysis identified four spa types among MRSA samples and differentiated MRSA strains accordingly. A nonsubjective differentiation model was developed according to genetic confidence percentage values produced by tested samples, which did not require visual interpretation of HRM curve results. The test was carried out at different settings, and result data were reanalyzed and confirmed with DNA sequencing. PCR-HRM curve analysis proved to be a robust and reliable test for spa typing and can be used as a tool in epidemiological studies.

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