Typing of Methicillin Resistant Staphylococcus Aureus Using DNA Fingerprints by Pulsed-field Gel Electrophoresis

Abstract

Background: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 analyze MRSA strains from the general population in Canton Sarajevo, B&H.Methods:Our investigation including either phenotypic and genotypic markers such as antimicrobial resistance, pulsed-field gel electrophoresis (PFGE), SCC typing, and Panton-Valentine leukocidin (PVL) detection.Results:Antimicrobial susceptibility: all MRSA isolates were resistant to the β-lactam antibiotics tested, and all isolates were susceptible trimethoprim sulphamethoxazole, rifampicin, fusidic acid, linezolid and vancomycin. Sixty-eight per cent of the MRSA isolates were resistant to erythromycin, 5% to clindamycin, 5% to gentamicin and 4% to ciprofloxacin. 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 (60%) and D: 27 (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.Conclusion:SCCmec type IV, together with the susceptibility profile and PFGE grouping, is considered to be typical of CA-MRSA

Livestock-Associated Methicillin-Resistant Staphylococcus aureus From Animals and Animal Products in the UK

“…DNA microarrays, which have been used for determining virulence or antimicrobial gene presence in diverse bacteria (Hopkins et al, 2007; Carter et al, 2008; Wragg et al, 2009), have also been applied to MRSA, as have WGS (Schouls et al, 2009; Monecke et al, 2011; Piccinini et al, 2012; Shore et al, 2012; Sharma et al, 2016; Sabat et al, 2017). The latter has been particularly useful and is increasingly being used instead of PFGE for purposes such as tracing outbreaks, as well as identifying the most likely source of acquisition, which is an essential component of an effective surveillance system to describe epidemiological trends and infection control strategies (Rebic et al, 2016; Sharma et al, 2016). In addition, although phenotypic methods are easier to perform and interpret, and they are cost effective and widely available, they may be less discriminatory.…”

“…In addition, although phenotypic methods are easier to perform and interpret, and they are cost effective and widely available, they may be less discriminatory. Genotypic methods, although more expensive and technically demanding, can provide more detailed characterisation of the isolate (Anjum, 2015; Rebic et al, 2016). Specifically, WGS is a more rapid method for identifying genetic determinants such as virulence and AMR genes, as well as studying phylogenetic relationships between groups of isolates based on the core genome of hundreds of genes, rather than DNA microarray.…”

Methicillin-resistant Staphylococcus aureus tracking spread among health-care workers and hospitalized patients in critical wards at a university hospital, Tehran, Iran

“…With the advent of molecular typing methods, epidemiologists can easily investigate outbreaks of infectious diseases, tracing the transmission of pathogens, and elucidate evolutionary relationships between microorganisms. Several genotyping methods such as amplified fragment length polymorphism [8] , [9] , multiple locus variable-number tandem repeat analysis [8] , [10] , [11] , multilocus sequence typing [12] , [13] , and pulsed-field gel electrophoresis (PFGE) [10] , [14] , [15] , [16] have been successfully applied for epidemiological investigations of MRSA infections worldwide. Of these methods, PFGE is still considered the reference standard fingerprinting method for monitoring and tracking MRSA dissemination in the hospital environment [17] .…”

The Dissemination and Molecular Characterization of Clonal Complex 361 (CC361) Methicillin-Resistant Staphylococcus aureus (MRSA) in Kuwait Hospitals

“…Molecular typing tools including staphylococcal protein A (spa) typing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis, staphylococcal cassette chromosome mec (SCC mec ) typing, DNA microarray, and whole genome sequencing (WGS) have been used in the epidemiologic surveillance of MRSA strains to detect and monitor emerging and reemerging infections as well as monitoring geographic spread and shifts of epidemic and endemic clones ( Pfaller, 1999 ; van Belkum et al, 2001 ; Monecke et al, 2011 ; Boswihi et al, 2016 , 2020a , 2020b ; Rebic et al, 2016 ). The application of these typing techniques to type MRSA from different geographic regions have shown that most of the MRSA infections reported worldwide were caused by a limited number of pandemic MRSA clones belonging to clonal complexes 5 (CC5), CC8/ST239, CC22, CC30, and CC45 ( Robinson and Enright, 2003 ; Monecke et al, 2011 ; Guthrie et al, 2020 ) although CA-MRSA isolates belong to more diverse genetic backgrounds compared to HA-MRSA ( Monecke et al, 2011 ; Udo, 2013 ; Tong et al, 2015 ; Guthrie et al, 2020 ).…”