Variable Number Tandem Repeat Fingerprinting Research Articles

Simultaneous carriage of multiple genotypes of Staphylococcus aureus in children

The co-existence of multiple genotypes in colonization by Staphylococcus aureus has not been fully investigated. The aim of this study was to evaluate the heterogeneity of S. aureus carriage in children. We evaluated 125 nasal and perianal swab samples that were positive for S. aureus from 76 children scheduled for elective surgery. For each sample, at least four colonies with the same or different morphotypes were selected for analysis. Multiple-locus variable-number tandem-repeat fingerprinting was used to determine the genetic relatedness and to characterize the clonality of the S. aureus strains. Of the 125 swabs, 91 (73 %) contained meticillin-sensitive S. aureus (MSSA), 8 (6 %) contained meticillin-resistant S. aureus (MRSA), and 26 (21 %) contained MSSA and MRSA simultaneously. A total of 738 S. aureus strains were evaluated with a mean of 6 colonies (range 4-15) picked from each culture. Of the 125 swabs, 32 (26 %) samples contained two genetically distinct S. aureus strains and 6 (5 %) contained three different genotypes. Multiple S. aureus strains simultaneously carried by individual children were genetically unrelated to each other. We concluded that the co-existence of multiple genotypes of S. aureus was common. The significance of multiple carriage is yet to be determined, but this intraspecies interplay could be important to pathogenicity and virulence in S. aureus.

Comparison of Two Multilocus Variable-Number Tandem-Repeat Methods and Pulsed-Field Gel Electrophoresis for Differentiating Highly Clonal Methicillin-Resistant Staphylococcus aureus Isolates

In the United Kingdom, EMRSA-15 and EMRSA-16 account for the majority (∼90%) of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infections. Currently, the standard typing technique, pulsed-field gel electrophoresis (PFGE), is laborious and insufficient for discriminating between closely related subtypes of EMRSA-15 and -16. The objective of the present study was to compare the usefulness of multilocus variable-number tandem-repeat fingerprinting (MLVF) and multilocus variable-number tandem-repeat analysis (MLVA) with PFGE for subtyping these highly clonal MRSA lineages. A panel of 85 MRSA isolates (41 EMRSA-15, 20 EMRSA-16, and 24 MRSA isolates with diverse PFGE patterns) was investigated. In addition, a further 29 EMRSA-15s with identical PFGE patterns from two geographically linked but epidemiologically distinct outbreaks and several sporadic cases were analyzed. PFGE, MLVF, and MLVA resolved 66 (Simpson's index of diversity [SID] = 0.984), 51 (SID = 0.95), and 42 (SID = 0.881) types, respectively, among the 85 MRSA isolates. MLVF was more discriminatory than MLVA for EMRSA-15 and -16 strains, but both methods had comparable discriminatory powers for distinguishing isolates in the group containing diverse PFGE types. MLVF was comparable to PFGE for resolving the EMRSA-15s but had a lower discriminatory power for the EMRSA-16s. MLVF and MLVA resolved the 29 isolates with identical PFGE patterns into seven and six subtypes, respectively. Importantly, both assays indicated that the two geographically related outbreaks were caused by distinct subtypes of EMRSA-15. Taken together, the data suggest that both methods are suitable for identifying and tracking specific subtypes of otherwise-indistinguishable MRSA. However, due to its greater discriminatory power, MLVF would be the most suitable alternative to PFGE for hospital outbreak investigations.

Prevalence and molecular characteristics ofStaphylococcus aureusisolates colonizing patients with atopic dermatitis and their close contacts in Singapore

Staphylococcus aureus colonization is an established pathogenic factor for disease flare in atopic dermatitis (AD). We conducted a study to investigate the colonization of S. aureus in patients with AD and their close contacts in order to evaluate the possibility of intrafamilial transmission. We sought to determine the distribution of the bacterial virulence factors and their correlation with disease severity. Nasal swabs and skin swabs (patients with AD only) were taken from patients with AD aged 2-21 years and their close contacts, seen at the National Skin Centre from January to March 2007. All S. aureus isolates were typed using multilocus variable-number tandem-repeat fingerprinting (MLVF) and screened for virulence factors via polymerase chain reaction (PCR) analysis. AD severity was determined by the SCORAD index. A total of 34 patients with AD and 55 close contacts were recruited. Thirty-one (91%) patients were colonized with S. aureus. Twenty-five (45%) of their close contacts were also colonized, and MLVF showed a high concordance of S. aureus isolates in index patients and their close contacts. On multivariate analysis, patients with a moderate SCORAD were more likely to be colonized by enterotoxin B-positive S. aureus (P = 0.027). No virulence factor was significantly associated with a severe SCORAD. The prevalence of S. aureus colonization was high among patients with AD and their close contacts. However, no predominant isolate of S. aureus was found to be associated with AD. The presence of superantigen B is possibly associated with moderate rather than severe disease in our population.

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

In this study, clonal relatedness of 202 Staphylococcus aureus (mostly methicillin-resistant) isolates recovered in 29 Polish hospitals was investigated by multiple-locus variable number tandem repeat fingerprinting (MLVF) and spa typing. Our analysis yielded 69 MLVF patterns and 34 spa types. Almost all isolates (97.4%) identical by MLVF were also indistinguishable by spa typing. Therefore, the MLVF method can be a cheap and fast screen before spa typing. Moreover, results obtained by MLVF suggest a set of simple criteria for grouping of spa types. The proposed algorithm groups isolates into the same cluster when spa sequences differ by a single mutation event: i) a single deletion or insertion of repeat unit(s) at the X region of the protein A gene or ii) a single nucleotide polymorphism within a repeat sequence. The combined use of these 2 methods, MLVF in local laboratories and spa typing of selected isolates in reference centers, can improve the monitoring of hospital-to-hospital strain transmission events and public health interventions on a huge scale.

Countrywide molecular survey of methicillin-resistant Staphylococcus aureus strains in Poland.

The present investigation was undertaken to assess the proportion of methicillin-resistant Staphylococcus aureus (MRSA) strains among hospital-acquired isolates and to determine the clones of MRSA currently circulating in Poland by using a number of molecular techniques. Between January and May 2005, methicillin resistance was investigated among a total of 915 S. aureus isolates collected from 39 hospitals. A total of 208 (22.7%) isolates were positive for the mecA gene by PCR. The molecular characterization of MRSA isolates was carried out by the multiple-locus variable-number tandem repeat fingerprinting, pulsed-field gel electrophoresis, multilocus sequence typing, and staphylococcal chromosomal cassette mec (SCCmec) typing methods. The Hungarian (PFGE B; ST239, SCCmec type III [ST239-III]), Iberian (ST247-I), and Berlin (ST45-IV) clones were predominant, representing approximately 52.9, 11.5, and 10.0% of the MRSA isolates, respectively. A decline in the proportion of earlier MRSA clones, such as ST5-IV (a Pediatric clone), ST80-IV) (a Mediterranean clone), ST239-III (a Polish and Brazilian clone), and ST30-IV (a southwest Pacific clone) was observed. Additionally, the emergence of an MRSA clone with SCCmec type V, possibly representing a community-acquired strain, was observed in two hospitals during this study.