Update to the Multiplex PCR Strategy for Assignment of
            <i>mec</i>
            Element Types in
            <i>Staphylococcus aureus</i>

Milheiriço, Catarina; Oliveira, Duarte C.; Lencastre, Herminia Garcez

doi:10.1128/aac.00275-07

Cited by 440 publications

(258 citation statements)

References 18 publications

(7 reference statements)

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“…The clones were classified based on the number of isolates and unique PFGE types as major (> 10 isolates per PFGE type), intermediate (4-9 isolates per PFGE type) or minor (2-3 isolates per PFGE types). In addition, spa typing and SCCmec typing were also performed (Harmsen et al, 2003;Milheirico et al, 2007). MLST was performed on a representative set of isolates from each major PFGE cluster (Enright et al, 2000).…”

Section: Methodsmentioning

confidence: 99%

Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage

Naicker

Karayem

Hoek

et al. 2016

Microbial Pathogenesis

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2 Declaration I, Preneshni Rochelle Naicker, hereby declare that the work contained in this assignment is my original work and that I have not previously submitted it, in its entirety or in part, at any university for a degree. The ability to form biofilms contributes significantly to the virulence of Staphylococcus aureus. Virulence factors may be associated with certain S. aureus lineages. However, the contribution of the genetic background of S. aureus to biofilm formation is poorly understood. This study investigated the association between the genetic background and the biofilm forming ability of clinical invasive S. aureus isolates. Secondary objectives included investigating any correlation with biofilm formation and methicillin resistance or the source of bacteraemia. Methods:The study was conducted at a 1300-bed tertiary hospital in Cape Town, South Africa. S. aureus isolates obtained from blood cultures between January 2010 and January 2012 were included. Genotypic characterization was performed by PFGE, spa typing, SCCmec typing and MLST. Thirty genotypically unique strains were assessed for phenotypic biofilm formation with the microtitre plate assay. All isolates were tested in triplicate and an average optical density, measured at a wavelength of 490nm, was determined. The biofilm forming ability of isolates with A 490 > 0.17 was considered 'weak positive' and A 490 > 0.34 'strong positive'. Isolates with A 490 ≤ 0.17 were considered non-adherent. ANOVA with Bonferroni-adjusted post-hoc tests and t-tests were used for statistical analysis of the association between biofilm forming ability and strain characteristics.Results: Fifty seven percent of isolates were capable of forming biofilms. Weak biofilm formation occurred in 40% (n=12) and strong biofilm formation in 17% (n=5) of isolates. Thirteen (43%) of the isolates were classified as non-adherent. All 5 isolates capable of strong biofilm formation belong to one spa clonal complex (spa-CC 064). Strains from spa-CC 064 were capable of higher biofilm formation than other spa clonal complexes (p=0.00002). These 5 strains belonged to MLST CC5 and CC8. Biofilm formation did not correlate with methicillin resistance and was not related to the source of bacteraemia. Conclusion:Biofilm formation correlates with the spa clonal lineage in our population of invasive S. aureus strains. High biofilm formation was associated with spa-CC 064. MLST CC5 and CC8 strains are capable of strong biofilm formation.

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Section: Methodsmentioning

confidence: 99%

Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage

Naicker

Karayem

Hoek

et al. 2016

Microbial Pathogenesis

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“…For all PCRs, 1 µl of lysate was used. Methicillin resistance was confirmed by PCR amplification of mecA DNA (Milheiriço et al, 2007a). Then, each isolate was submitted to SCCmec typing and SCCmec type IV subtyping methods.…”

Section: Methodsmentioning

confidence: 99%

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Djoudi

Bonura

Touati

et al. 2016

Journal of Medical Microbiology

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Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcus haemolyticus, 7 Staphylococcus aureus, 4 Staphylococcus sciuri, 2 Staphylococcus saprophyticus and 1 isolate each of Staphylococcus xylosus and Staphylococcus lentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new b-lactam resistance.

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“…9 Genotyping of the MRSA isolates was routinely performed by multilocus variable number tandem repeat fingerprinting (MLVF). 10 Banding patterns were analyzed both visually and by using Bionumerics version 5.10 (Applied Maths, Sint-Martens-Latem, Belgium).…”

Section: Molecular Typing Of Mrsa Isolatesmentioning

confidence: 99%

Methicillin-resistant Staphylococcus aureus nasal colonization in a level III neonatal intensive care unit: Incidence and risk factors

Giuffrè¹,

Amodio²,

Bonura³

et al. 2015

American Journal of Infection Control

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Update to the Multiplex PCR Strategy for Assignment of mec Element Types in Staphylococcus aureus

Cited by 440 publications

References 18 publications

Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage

Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Methicillin-resistant Staphylococcus aureus nasal colonization in a level III neonatal intensive care unit: Incidence and risk factors

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