Staphylococcus aureus es un patógeno capaz de causar infecciones con amplio rango de severidad y adaptarse a diferentes tejidos. Su epidemiología es compleja, por circulación de cientos de clones a nivel mundial, lo que requiere de métodos moleculares reproducibles y de alto poder discriminatorio para la identificación de los mismos. El presente estudio tuvo como objetivo principal la estandarización del análisis multi-locus de número variable de repeticiones en tándem (MLVA) para análisis de variabilidad genética de aislados de S. aureus previamente tipificados por electroforesis en gel de campo pulsado (PFGE), gold standard para tipificación de aislados. La MLVA se realizó por amplificación de 7 locus VNTR (clfA, clfB, sdrC, sdrD, sdrE, sspA y spA) por PCR. Se alcanzó un alto nivel de reproducibilidad. El empleo de cepas previamente tipificadas por análisis de secuencias multi-locus (MLST), PFGE, locus spa y cassette SCCmec, permitió validar de forma comparativa el agrupamiento generado por MLVA. Los aislados que fueron agrupados como idénticos por MLVA presentaron resultados congruentes con la totalidad de las otras técnicas moleculares y ésta demostró ser más sensible que PFGE para distinguir entre aislados que presentaron patrones PFGE idénticos. La MLVA cumple todos los criterios de un método de tipificación útil.
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is one of the first causes of skin and soft tissue infections, and can also produce severe diseases such as osteomyelitis and pneumonia. The aim of this descriptive study was to determine the SCCmec type and virulence profile and to study the genetic diversity by MLVA analysis of 21 CA-MRSA isolates that infected Paraguayan children in 2010. The SCCmec type and virulence factors were performed by PCR and genetic diversity by MLVA (multiple locus variable analysis). All the isolates carried SCCmec cassette iv. hla, hlb and sea genes were detected in 28,6%, 9,5% and 4,8% respectively. The MLVA analysis showed high genetic diversity with congruent antibiotic resistance and virulence profiles. This study revealed the presence of CA-MRSA harbouring SCCmeciv with high genetic diversity, providing information not available in our country.
Background: Patients undergoing hemodialysis are susceptible to the nasal carriage of Staphylococcus aureus, increasing the risk of developing infections associated with higher morbidity and mortality. The objective of this study was to describe the frequency of S. aureus carriage in hemodialysis patients and to perform molecular analysis of isolates by applying multiple-locus variable analysis. Methods: We conducted a descriptive cross sectional study with non-probabilistic sampling that included 28 hemodialysis patients attending the Nephrology Department of Hospital de Clínicas in Asunción, Paraguay. We obtained clinical data from medical records and interviews with patients. Nasal swabs were collected and analyzed by microbiological and molecular methods. Results: The frequency of S. aureus carriage was 50% (14/28), 93% of which (13/14) were methicillin resistant, 57% (6/14) were gentamicin resistant and 36% (5/14) were resistant to more than 4 antibiotic classes. S. aureus carriers showed higher frequency of rhinitis (p=0.02 odds ratio [OR]=6.6 (1.2- 34.4)). Seven methicillin-resistant S. aureus isolates had been analyzed by multiple-locus variable analysis, two of them showed identical pattern bands. Conclusion: We found a high frequency of methicillin-resistant Staphylococcus aureus colonization and the presence of two isolates with identical profile in the multiple-locus variable analysis indicating the possibility of transmission between patients.
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major human pathogens, causing a wide range of infections from food poisoning to necrotizing pneumonia, endocarditis, or septicemia. It could carry numerous resistance genes and virulence factors, some of which are related to the severity of the infection, being also regarded as a potential "Super Bug". In Paraguay, the prevalence of CC30-ST30-IV clones is the leading cause of S. aureus infections both at the regional level and in pediatric population. Here we aimed to deeply analyze the genomic features of MRSA isolates that cause invasive infections in Paraguayan children. An observational, descriptive, cross-sectional study was designed to analyze representative MRSA isolates of the main clones identified between 2009 to 2013 in Paraguayan children. All the genetically linked MRSA isolates were recovered from diverse clinical sources, patients, and hospitals at broad gap periods. Cases were primarily community-acquired, which excludes in-hospital transmission or outbreaks. MRSA isolates were analyzed with short-read paired-end sequencing and assessed for the virulome, resistome, and phylogenetic relationships. The pan-genomic analysis of these clones revealed three major and different clonal complexes (CC8, CC30, and CC5), each composed of clones closely related to each other, despite having different spa types. Furthermore, multiple virulence and resistance genes were identified for the first time in this study, pointing out the complex virulence profiles of MRSA circulating in the country. This study opens a wide range of new possibilities for future projects and trials to improve the existing knowledge on the epidemiology of MRSA circulating in Paraguay.
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