Molecular characterization of methicillin-resistant Panton-valentine leukocidin positive staphylococcus aureus clones disseminating in Tunisian hospitals and in the community (original) (raw)
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2020
Background: The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. Methods: A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antibiotic resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multi-locus sequence type and SCC mec typing. Results: Of 146 Staphylococcus aureus isolates 24 (16.4%) isolates identified as MRSA strains. According to antibiotic susceptibility testing the highest resistance rates were seen to erythromycin, cefoxitin and clindamycin. All of Staphylococcus aureus isolates were sensitive to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCC mec types were obtained among MRSA strains including 16 (66.7%) SCC mec type V, 3 (12.5%) SCC mec type III and 5 (20.8%) SCC mec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%) , cna (25.0%), fnbB (16.6%) , ebps 5 (20.8%), and fnbA , bbp and clfA genes were not detected. MLST analysis revealed 11 sequence types among MRSA isolates as follows:
Microbiologia Medica, 2019
Resistance to methicillin in methicillin resistant Staphylococcus aureus (MRSA) is dependent on mecA gene located on staphylococcal cassette chromosome (SCC). Both SCCmec type and Panton-Valentine leukocidin (PVL) affect S. aureus pathogenicity. Aim of this study was to investigate the prevalence of SCCmecA types and pvl genes among MRSA isolates from inpatients. During this cross-sectional study on 100 clinical isolates, following antibiotic susceptibility test, screening of mecA and pvl genes, as well as SCCmec typing, was done in a multiplex PCR technique. From the studied samples, 58 isolates were recognized as MRSA. The frequency of mecA and pvl was 58% and 4%, respectively. All of the MRSA were resistant to cefoxitin and had the highest sensitivity to chloramphenicol. The majority (77.5%) of MRSA was originated from wound samples. The SCCmec III was the most frequent type (22.4%) in these samples. The pvl positive isolates were from SCCmec IVb and V, thus meaning they are from...
Antimicrobial Resistance and Infection Control, 2020
Background: The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. Methods: A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antimicrobial resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multilocus sequence typing and SCCmec typing. Results: Of 146 Staphylococcus aureus isolates, 24 (16.4%) isolates were identified as MRSA strains. According to antimicrobial susceptibility testing the highest resistance rates were seen for tetracycline, erythromycin, ciprofloxacin and gentamicin. All of Staphylococcus aureus isolates were susceptible to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCCmec types were obtained among MRSA strains including 16 (66.7%) SCCmec type V, 3 (12.5%) SCCmec type III and 5 (20.8%) SCCmec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%), cna (25.0%), fnbB (16.6%), ebps 5 (20.8%), and the fnbA, bbp and clfA genes were not detected in any MRSA isolate. MLST analysis revealed 11 sequence types among MRSA isolates as follows:
Journal of Medical Microbiology and Infectious Diseases, 2019
Introduction: Staphylococcus aureus is among the primary cause of hospitals and community-acquired infections. The emergence of methicillin-resistant S. aureus (MRSA) strains has resulted in the treatment failure of the infections caused by these bacteria. Hence, regional data on antibiotic resistance of S. aureus strains is necessary to adopt appropriate treatment regimens. This study aims to identify the diversities and their frequencies among MRSA isolates by molecular analysis of four genes. Methods: In a cross-sectional study, 100 S. aureus isolates from patients hospitalized in two hospitals of Ahvaz, Iran were collected and identified. The MRSA isolates were identified by phenotypic method and amplification of the mecA gene. The diversity of MRSA isolates was investigated by amplification of the coa, spa, aroA, and gap genes followed by RFLP analysis using the AluI, HindIII, TaqI and RsaI restriction enzymes. Results: In this study, we identified 50 MRSA isolates. Based on the analysis of coa gene, 8 types, spa gene 5 types and 17 subtypes, coa gene with AluI 13 types, and spa with HindIII 13 types were identified. Also, the RFLP analysis of gap gene with AluI revealed 3 types, and of aroA gene with TaqI and RsaI, 3 types and 2 subtypes, respectively. Conclusion: Our PCR-RFLP analysis revealed that diversities are present among MRSA isolates originated from clinical samples and showed that this method is simple, reproducible, and cost-effective.
Archives of Microbiology, 2010
Staphylococcus aureus is known as an invasive human pathogen, resulting in significant morbidity and mortality worldwide; however, information on community-associated S. aureus (CA-SA) from bloodstream infections (BSI) in children in China remains scarce. This study aimed to investigate the molecular characteristics of 78 CA-SA isolates recovered from pediatric patients with BSI between 2012 and 2017 in Shanghai. All isolates including 51 (65.4%) methicillin-susceptible S. aureus (MSSA) and 27 (34.6%) methicillin-resistant S. aureus (MRSA) isolates were characterized based on antimicrobial resistance, virulence genes, multilocus sequence typing (MLST), spa, and SCCmec typing. A total of 18 distinct sequence types (STs) and 44 spa types were identified. ST188 and ST7 were the predominant MSSA clones and ST59-MRSA-SCCmecIV/V was the most common MRSA clone. Spa t189 (9.0%, 7/78) was the most common spa type. SCCmec types IV and V were observed at frequencies of 59.3 and 40.7%, respectively. Notably, 40 (51.3%) S. aureus BSI strains were multidrug resistant (MDR), and these were mostly resistant to penicillin, erythromycin, and clindamycin. MRSA strains were associated with substantially higher rates of resistance to multiple antibiotics than MSSA strains. Fifty (64.1%, 50/78) isolates, including 19 (70.3%) MRSA isolates, harbored ≥ 10 tested virulence genes, as evaluated in this study. Ten (37.0%) MRSA isolates and four (7.8%) MSSA isolates harbored the gene encoding Panton-Valentine leukocidin (PVL). Virulence genes analysis showed diversity in different clones; the seb-sek-seq genes were present in all ST59 strains, whereas the seg-sei-sem-senseo genes were present in different clones including ST5, ST20, ST22, ST25, ST26, ST30, ST121, and ST487 strains. In conclusion, this study revealed that communityassociated S. aureus strains from BSI in children demonstrated considerable genetic diversity, and identified major genotypes of CA-MRSA and CA-MSSA, with a high
Diagnostic Microbiology and Infectious Disease, 2013
The spread of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) has been reported in communities worldwide. In this study, we characterized 64 Tunisian CA-MRSA by agr typing, polymerase chain reaction assay for 20 virulence genes, staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and protein A gene (spa) typing. All our isolates were lukS-PV-lukF-PV positive, etd positive, and edin positive. They harbored SCCmec type IV and belonged to agr group 3. PFGE typing showed that our isolates were distributed in 11 different pulsotypes. spa typing and MLST, performed with isolates representative of each PFGE pattern, revealed that all isolates had a unique spa type (t044) and a common sequence type (ST80). The isolates showed susceptibility to the majority of antibiotics, and resistance to kanamycin, erythromycin, and tetracycline, but intermediate resistance to fusidic acid. Full analysis of our results revealed that our isolates were nonmultiresistant and belonged to a single clonal type ST80.
Central European Journal of Public Health
Objectives: Staphylococcus aureus (SA) represents one of the most important microorganism that is part of the normal microflora of humans, but in certain conditions can cause very serious infections. Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for a wide spectrum of nosocomial and community associated infections worldwide. The aim of this study was to determine community acquired MRSA (CA-MRSA), as well as the frequency of Panton-Valentine leukocidin (PVL) genes and staphylococcal cassette chromosome mec (SCCmec) types in isolates obtained from outpatients in the region of 700,000 people (Canton Sarajevo, Bosnia and Herzegovina) Methods: Our investigation included phenotypic and genotypic markers such as antimicrobial resistance, pulsed-field gel electrophoresis (PFGE), SCC typing, and PVL detection. Results: Antimicrobial susceptibility: all MRSA isolates were resistant to the β-lactam antibiotics tested, and all isolates were susceptible to trimethoprim sulphamethoxazole, rifampicin, fusidic acid, linezolid, and vancomycin. After the PFGE analysis, the isolates were grouped into five similarity groups: A-E. The largest number of isolates belonged to one of two groups: C-60% and D-27%. In both groups C and D, SCCmec type IV was predominant (60% and 88.8%, respectively). A total of 24% of the isolates had positive expression of PVL genes, while 76% showed a statistically significantly greater negative expression of PVL genes. Conclusions: Using combination techniques, we were able to investigate the origin and genetic background of the strains. PFGE analysis revealed two large, genetically related groups of strains consisting of 87 isolates. Our results suggest failure to apply the screening policy, and a lack of knowledge about multiresistant MRSA strains. This study showed the local epidemiological situation which should be the basis of antimicrobial empiric therapy for non-hospitalized patients.
Journal of Clinical Microbiology, 2007
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) carrying pvl is an emerging problem worldwide. CA-MRSA tends to harbor staphylococcal cassette chromosome mec type IV (SCC mec IV), to be non-multiantibiotic resistant, and to have different genotypes from the local hospital-acquired MRSA (HA-MRSA). However, in Ireland, 80% of HA-MRSA isolates have the non-multiantibiotic-resistant genotype ST22-MRSA-IV. This study investigated MRSA isolates from Ireland (CA-MRSA, health care-associated MRSA, and HA-MRSA) for the carriage of pvl and determined the genotypic characteristics of all pvl -positive isolates identified. All 1,389 MRSA isolates were investigated by antibiogram-resistogram typing and SmaI DNA macrorestriction analysis. pvl -positive isolates were further characterized by multilocus sequence typing and SCC mec , agr , and toxin gene typing. Twenty-five (1.8%) MRSA isolates belonging to six genotypes (ST30, ST8, ST22, ST80, ST5, and ST154) harbored pvl ....
Jundishapur Journal of Microbiology, 2015
Background: Staphylococcus aureus, an important human pathogen is one of the main causative agents of nosocomial infection. Virulence genes play a major role in the pathogenicity of this agent and its infections. Methicillin-Resistant Staphylococcus aureus (MRSA) isolates are major challenge among infectious agents that can cause severe infections and mortality. Methicillin-resistant S. aureus produces a unique type of Penicillin Binding Protein 2a (PBP2a) that has low affinity for β-lactam antibiotics. Most of the MRSA bacterial strains can also produce a leukotoxin as Panton-Valentine Leukocidin (PVL) that increases the virulence of MRSA strains and can cause severe necrotic pneumonia. The presence of pvl gene is a genetic marker for the MRSA populations. Objectives: The aim of this study was to explore the association of pvl and mecA genes in clinical isolates of MRSA. Materials and Methods: Fifty MRSA isolates were collected from 200 clinical samples from three different educational hospitals in Ahvaz, Iran, and identified by biochemical tests including catalase, oxidase, tube coagulase, mannitol fermentation, and sensitivity to furazolidone, resistance to bacitracin, PYR test and Voges-Proskauer test. Their resistance to methicillin was evaluated using the disc diffusion method. DNA was extracted by boiling and then the presence of pvl and mecA genes was investigated by the polymerase chain reaction method using specific primers. Results: The results revealed that mecA and pvl genes were positive for 15 (30%) and 3 (6%) of the isolates, respectively. None of mecA positive isolates was positive for pvl gene. Conclusions: It can be concluded from these results that fortunately the prevalence of pvl gene is low in MRSA isolates in this region and there is no association between the presence of pvl and mecA genes in these isolates.