title: Design of two molecular methodologies for the rapid identification of Colombian community-acquired methicillin-resistant Staphylococcus aureus isolates.
abstract: BACKGROUND
  Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections are found with increasing the frequency, both in healthy individuals in the community and in hospitalized patients. In Colombia and the Andean region, CA-MRSA isolates have a genetic background that is related to the pandemic USA300 clone.
  
  
  OBJECTIVE
  Two molecular methods are designed and standardized for the rapid differentiation of Colombian community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) isolates.
  
  
  METHODS
  Two molecular methods were standardized for the identification of CA-MRSA isolates. The first method was based on the differential digestion of the carbamate kinase (arcC)and guanylate kinase (gmk) genes in the sequences type 5 (ST5) in the HA-MRSA isolates and 8 (ST8) in the CA-MRSA isolates. The second method was based on the PCR amplification of 5 specific virulence factors found in CA-MRSA and HA-MRSA isolates. The specificity and precision of each method were evaluated using 237 clinical MRSA isolates.
  
  
  RESULTS
  The first method identified 100% and 93.2% of the CA-MRSA and HA-MRSA isolates, respectively. The second method also correctly identified the two isolates types (CA-MRSA and HA-MRSA).
  
  
  CONCLUSIONS
  These two methods are a convenient alternative for the rapid identification of the CA-MRSA isolates, compared with other techniques such as pulsed field gel electrophoresis and multilocus sequence typing, which are time-consuming and more expensive.
authors.affiliation: Laboratorio de Genética Molecular Bacteriana, Universidad El Bosque, Bogotá, DC, Colombia. javiesco21@yahoo.com
authors.forename: Javier Antonio
authors.initials: JA
authors.lastname: Escobar
authors.forename: Ingrid Tatiana
authors.initials: IT
authors.lastname: Gómez
authors.forename: Martha Johanna
authors.initials: MJ
authors.lastname: Murillo
authors.forename: Betsy Esperanza
authors.initials: BE
authors.lastname: Castro
authors.forename: Bibiana
authors.initials: B
authors.lastname: Chavarro
authors.forename: Ricaurte Alejandro
authors.initials: RA
authors.lastname: Márquez
authors.forename: Natasha
authors.initials: N
authors.lastname: Vanegas
mesh_terms: D000483:Alleles; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods* / Q000592:standards; D003105:Colombia; D017714:Community-Acquired Infections / Q000453:epidemiology / Q000382:microbiology; D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology; D016521:Electrophoresis, Gel, Pulsed-Field; D005798:Genes, Bacterial; D051528:Guanylate Kinases / Q000235:genetics; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology*; D013997:Time Factors; D014774:Virulence / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D017851:Phosphotransferases (Carboxyl Group Acceptor); C019288:carbamate kinase; D051528:Guanylate Kinases