Comparison of Fingerprinting Methods for Typing Methicillin-Resistant Staphylococcus aureus Sequence Type 398

ABSTRACT This study evaluates the multiple-locus variable-number tandem-repeat assay (MLVA) and pulsed-field gel electrophoresis (PFGE) when using restriction enzymes BstZI, SacII, and ApaI to fingerprint a diverse collection of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) sequence type 398 (ST398) isolates. These isolates had been characterized previously by multilocus sequence typing, spa typing, and staphylococcal cassette chromosome mec (SCCmec) typing. Typeability and discriminatory power were analyzed, and the concordance between the various methods was determined. All MRSA ST398 isolates were typeable by the MLVA and PFGE using BstZI, SacII, and ApaI. With each method, the MRSA ST398 isolates formed a separate group from the two non-ST398 MRSA strains. PFGE, performed with any of the three restriction enzymes, had the most discriminatory power, followed by MLVA, spa typing, and SCCmec typing. The MLVA showed the highest concordance with PFGE using ApaI and spa typing. As further expressed by the Wallace coefficient, the MLVA type was poorly predicted by spa typing, whereas the spa type was well predicted by MLVA. PFGE, using a combination of all three restriction enzymes, had the highest concordance with the MLVA but had a low probability of being predicted by MLVA. PFGE, using a combination of all three restriction enzymes, was able to predict SCCmec type and MLVA type completely and had a high probability of predicting spa type. Both the MLVA and PFGE could be used to discriminate among the MRSA ST398 isolates. Although the MLVA is a faster technique, PFGE had more discriminatory power than the MLVA, especially when a combination of restriction enzymes was used.

[1]  C. Suetens,et al.  Methicillin-Resistant Staphylococcus aureus ST398 in Swine Farm Personnel, Belgium , 2009, Emerging infectious diseases.

[2]  W. Hryniewicz,et al.  Molecular surveillance of methicillin-resistant Staphylococcus aureus by multiple-locus variable number tandem repeat fingerprinting (formerly multiple-locus variable number tandem repeat analysis) and spa typing in a hierarchic approach. , 2008, Diagnostic microbiology and infectious disease.

[3]  J. Krzysztoń-Russjan,et al.  Usefulness of Multiple-Locus VNTR Fingerprinting in detection of clonality of community- and hospital-acquired Staphylococcus aureus isolates , 2008, Antonie van Leeuwenhoek.

[4]  A. Voss,et al.  MRSA in livestock animals-an epidemic waiting to happen? , 2008, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[5]  A. van Belkum,et al.  Methicillin-Resistant and -Susceptible Staphylococcus aureus Sequence Type 398 in Pigs and Humans , 2008, Emerging infectious diseases.

[6]  Andreas Voss,et al.  Emergence of Methicillin-Resistant Staphylococcus aureus of Animal Origin in Humans , 2007, Emerging infectious diseases.

[7]  H. de Lencastre,et al.  Multiplex PCR strategy for subtyping the staphylococcal cassette chromosome mec type IV in methicillin-resistant Staphylococcus aureus: 'SCCmec IV multiplex'. , 2007, The Journal of antimicrobial chemotherapy.

[8]  H. Boshuizen,et al.  High prevalence of methicillin resistant Staphylococcus aureus in pigs. , 2007, Veterinary microbiology.

[9]  J. Mcgowan,et al.  Multiple-Locus Variable-Number Tandem-Repeat Assay Analysis of Methicillin-Resistant Staphylococcus aureus Strains , 2007, Journal of Clinical Microbiology.

[10]  W. Hryniewicz,et al.  Comparison of PCR-Based Methods for Typing Staphylococcus aureus Isolates , 2006, Journal of Clinical Microbiology.

[11]  J A Carriço,et al.  Illustration of a Common Framework for Relating Multiple Typing Methods by Application to Macrolide-Resistant Streptococcus pyogenes , 2006, Journal of Clinical Microbiology.

[12]  A. Voss,et al.  Presence of a Novel DNA Methylation Enzyme in Methicillin-Resistant Staphylococcus aureus Isolates Associated with Pig Farming Leads to Uninterpretable Results in Standard Pulsed-Field Gel Electrophoresis Analysis , 2006, Journal of Clinical Microbiology.

[13]  A. van Belkum,et al.  Distribution of the Serine-Aspartate Repeat Protein-Encoding sdr Genes among Nasal-Carriage and Invasive Staphylococcus aureus Strains , 2006, Journal of Clinical Microbiology.

[14]  B. Oppenheim,et al.  Use of Variations in Staphylococcal Interspersed Repeat Units for Molecular Typing of Methicillin-Resistant Staphylococcus aureus Strains , 2006, Journal of Clinical Microbiology.

[15]  A. Voss,et al.  Methicillin-resistant Staphylococcus aureus in Pig Farming , 2005, Emerging infectious diseases.

[16]  T. Louie,et al.  Novel Multiplex PCR Assay for Characterization and Concomitant Subtyping of Staphylococcal Cassette Chromosome mec Types I to V in Methicillin-Resistant Staphylococcus aureus , 2005, Journal of Clinical Microbiology.

[17]  J. Krzysztoń-Russjan,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 , 2005, Journal of Clinical Microbiology.

[18]  P. François,et al.  Use of an Automated Multiple-Locus, Variable-Number Tandem Repeat-Based Method for Rapid and High-Throughput Genotyping of Staphylococcus aureus Isolates , 2005, Journal of Clinical Microbiology.

[19]  D. Ussery,et al.  Distribution and characterization of staphylococcal interspersed repeat units (SIRUs) and potential use for strain differentiation. , 2004, Microbiology.

[20]  Javier Garaizar,et al.  In silico analysis of complete bacterial genomes: PCR, AFLP-PCR and endonuclease restriction , 2004, Bioinform..

[21]  Susan K. Johnson,et al.  Comparison of community- and health care-associated methicillin-resistant Staphylococcus aureus infection. , 2003, JAMA.

[22]  M. Kaufmann,et al.  Harmonization of Pulsed-Field Gel Electrophoresis Protocols for Epidemiological Typing of Strains of Methicillin-Resistant Staphylococcus aureus: a Single Approach Developed by Consensus in 10 European Laboratories and Its Application for Tracing the Spread of Related Strains , 2003, Journal of Clinical Microbiology.

[23]  James Travis,et al.  New Method for Typing Staphylococcus aureus Strains: Multiple-Locus Variable-Number Tandem Repeat Analysis of Polymorphism and Genetic Relationships of Clinical Isolates , 2003, Journal of Clinical Microbiology.

[24]  D. Oliveira,et al.  Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus , 2002, Antimicrobial Agents and Chemotherapy.

[25]  M. Struelens,et al.  Epidemiologic typing and delineation of genetic relatedness of methicillin-resistant Staphylococcus aureus by macrorestriction analysis of genomic DNA by using pulsed-field gel electrophoresis , 1992, Journal of clinical microbiology.

[26]  P. Hunter Reproducibility and indices of discriminatory power of microbial typing methods , 1990, Journal of clinical microbiology.

[27]  P. Hunter,et al.  Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity , 1988, Journal of clinical microbiology.

[28]  M. Thomashow,et al.  Introduction of pAM beta 1 into Listeria monocytogenes by conjugation and homology between native L. monocytogenes plasmids , 1984, Infection and immunity.

[29]  M. Finland,et al.  Methicillin-resistant Staphylococcus aureus at Boston City Hospital. Bacteriologic and epidemiologic observations. , 1968, The New England journal of medicine.

[30]  B. Spratt,et al.  Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. , 2000, Journal of clinical microbiology.