Keywords
epidemiology
bacterial typing techniques
molecular typing.
How to Cite
Abstract
Staphylococcus aureus is a pathogen that can produce several infections with a wide range of severity and it has the hability to adapt to different tissues. The epidemiology is complex, by circulation of many differents clones worlwide, so the analysis for its identification requires reproducible and high discriminatory powder molecular methods. The aim of this study was to standardize the molecular technique multiple-locus variable number of tandem repeat analysis (MLVA) for the genetic variability analysis of S. aureus isolates, previously characterized by pulsed field gel electrophoresis (PFGE). The MLVA was made by PCR amplification of seven VNTR locus (clfA, clfB, sdrC, sdrD, sdrE, sspA y spA). A high level of reproducibility has reached in the study. The use of isolates previously typified by multi-locus secuencing typing (MLST), PFGE, locus spa and cassette SCCmec, allowed to validate the MLVA clusters comparatively. The isolates that were clustered by MLVA as the same isolate, showed the same results by other molecular techniques, and the MLVA can distinguish isolates with identical PFGE patterns. This technique have all criteria of a usefull molecular typification technique.References
Collery MM, Smyth DS, Twohig JM, Shore AC, Coleman DC, Smyth CJ. Molecular Typing of Nasal Carriage Isolates of Staphylococcus aureus from an Irish University Student Population Based on Toxin Gene PCR, Agr Locus Types and Multiple Locus, Variable Number Tandem Repeat Analysis. J Med Microbiol. 2008;57(3):348-58.
Paganini M, Della LP, Muller OB, Ezcurra G, Uranga M, Aguirre C, et al. Infecciones por Staphylococcus aureus resistente a meticilina adquiridas en la comunidad en niños antes sanos y en niños relacionados al hospital en la Argentina. Rev Chil Infectol. 2009;26(5):406-12.
Pertuz-Meza Y, Perez-Quintero C, Pabón-Varela Y. Aspectos epidemiológicos de la sepsis, en unidades de cuidados intensivos Santa Marta, Colombia. Duazary. 2016;13(2):126-32.
Schouls LM, Spalburg EC, van Luit M, Huijsdens XW, Pluister GN, van Santen-Verheuvel MG, et al. Multiple-Locus Variable Number Tandem Repeat Analysis of Staphylococcus aureus: Comparison with Pulsed-Field Gel Electrophoresis and spa-Typing. PLoS ONE. 2009;4(4).
Malachowa N, Sabat A, Gniadkowski M, Krzyszton-Russjan J, Empel J, Miedzobrodzki J, et al. Comparison of Multiple-Locus Variable-Number Tandem-Repeat Analysis with Pulsed-Field Gel Electrophoresis, spa Typing, and Multilocus Sequence Typing for Clonal Characterization of Staphylococcus aureus Isolates. J Clin Microbiol. 2005;43(7):3095-100.
Sabat A, Krzyszton-Russjan J, Strzalka W, Filipek R, Kosowska K, Hryniewicz W, et al. New Method for Typing Staphylococcus aureus Strains: Multiple-Locus Variable-Number Tandem Repeat Analysis of Polymorphism and Genetic Relationships of Clinical Isolates. J Clin Microbiol. 2003;41(4):1801-4.
Rivero-Pérez B, Pérez-Roth E, Méndez-Álvarez S. Evaluation of Multiple-Locus Variable-Number Tandem-Repeat Analysis for Typing a Polyclonal Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Population in an Area Where Such Infections Are Endemic. J Clin Microbiol. 2010;48(8):2991-4.
Pérez-Roth E, Claverie-Martín F, Villar J, Méndez-Álvarez S. Multiplex PCR for Simultaneous Identification of Staphylococcus aureus and Detection of Methicillin and Mupirocin Resistance. J Clin Microbiol. 2001;39(11):4037-41.
Mayor L, Ortellado J, Menacho C, Lird G, Courtier C, Gardon C, et al. Molecular Characterization of Methicillin-Resistant Staphylococcus aureus Isolates Collected in Asunción, Paraguay. J Clin Microbiol. 2007;45(7):2298-300.
Weiler, N, Leotta, GA, Zarate, MN, Manfredi, E, Alvarez, ME, Rivas, M. Brote de intoxicación alimentaria asociado al consumo de leche ultrapasteurizada en la República del Paraguay. Revista Argentina de Microbiología. 2011;43:33-6.
Rodríguez F, Carpinelli L, Basualdo W, Castro H, Quiñónez B, Arguello R, et al. Frecuencia de genes que codifican factores de virulencia en Staphylococcus aureus aislados de niños que concurrieron al Hospital General Pediátrico Niños de Acosta Ñú, durante el año 2010. Mem IICS. 2015;13(1):58-66.
Shopsin B, Gomez M, Montgomery SO, Smith DH, Waddington M, Dodge DE, et al. Evaluation of Protein A Gene Polymorphic Region DNA Sequencing for Typing of Staphylococcus aureus Strains. J Clin Microbiol. 1999;37(11):3556-63.
Kondo Y, Ito T, Ma XX, Watanabe S, Kreiswirth BN, Etienne J, et al. Combination of Multiplex PCRs for Staphylococcal Cassette Chromosome mec Type Assignment: Rapid Identification System for mec, ccr, and Major Differences in Junkyard Regions. Antimicrob Agents Chemother. 2007;51(1):264-74.
Enright MC, Day NPJ, Davies CE, Peacock SJ, Spratt BG. Multilocus Sequence Typing for Characterization of Methicillin-Resistant and Methicillin-Susceptible Clones of Staphylococcus aureus. J Clin Microbiol. 2000;38(3):1008-15.
Chung M, de Lencastre H, Matthews P, Tomasz A, Adamsson I, Aires de Sousa M, et al. Molecular typing of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis: comparison of results obtained in a multilaboratory effort using identical protocols and MRSA strains. Microb Drug Resist Larchmt N. 2000;6(3):189-98.
Zhang K, Sparling J, Chow BL, Elsayed S, Hussain Z, Church DL, et al. New quadriplex PCR assay for detection of methicillin and mupirocin resistance and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci. J Clin Microbiol. 2004;42(11):4947-55.
Sambrook J, Fritsch EF. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory; 1989.
Holmes A, Edwards GF, Girvan EK, Hannant W, Danial J, Fitzgerald JR, et al. Comparison of Two Multilocus Variable-Number Tandem-Repeat Methods and Pulsed-Field Gel Electrophoresis for Differentiating Highly Clonal Methicillin-Resistant Staphylococcus aureus Isolates. J Clin Microbiol. 2010;48(10):3600-7.
Tenover FC, Vaughn RR, McDougal LK, Fosheim GE, McGowan JE. Multiple-Locus Variable-Number Tandem-Repeat Assay Analysis of Methicillin-Resistant Staphylococcus aureus Strains. J Clin Microbiol. 2007;45(7):2215-9.
Moser SA, Box MJ, Patel M, Amaya M, Schelonka R, Waites KB. Multiple-locus variable-number tandem-repeat analysis of meticillin-resistant Staphylococcus aureus discriminates within USA pulsed-field gel electrophoresis types. J Hosp Infect. 2009;71(4):333-9.
Bustamante AV, Lucchesi PMA, Parma AE. Molecular characterization of Verocytotoxigenic Escherichia coli O157:H7 isolates from Argentina by Multiple-Loci VNTR Analysis (MLVA). Braz J Microbiol. 2009;40(4):927-32.
Blomfeldt A, Hasan AA, Aamot HV. Can MLVA Differentiate among Endemic-Like MRSA Isolates with Identical Spa-Type in a Low-Prevalence Region? PloS One. 2016;11(2):e0148772.