Background Outbreaks of Panton-Valentine Leucocidin (PVL)-producing methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections (SSTIs) are a recurrent challenge for the Royal Marines at the Commando Training Centre (CTCRM). The intensity of commando training, its impact on skin integrity, and persistent colonisation reservoirs within the training centre have thwarted attempts to prevent these outbreaks. Aim To present an outbreak of PVL-producing MRSA SSTIs at a military training centre, demonstrating the benefit of additional abrasion and laceration swabs on the identification of colonised personnel and showing the effectiveness of a 10-day decolonisation regime. Method Following the identification of the outbreak of PVL-producing MRSA, all 36 members of the Recruit Troop underwent nasal MRSA screening to identify MRSA carriers. The screening was repeated on day 16 after completing an enhanced 10-day decolonisation regime. A third screening was conducted on the 110th day after a second peak of infection was identified. Various infection control measures, such as enhanced cleaning, restriction of movement and adjustments to the military training serials, were introduced to prevent further spread through the training centre. Results In this outbreak, two-thirds (eighteen) of the Recruit Troop suffered MRSA-PVL skin infections requiring antibiotic therapy and three required hospital admission for surgical management of their abscesses. The outbreak lasted 130 days, with two spikes in infections 10 weeks apart. The outbreak was successfully confined to one troop. Conclusion With concerns about low identification rates of carriers using nasal screening for MRSA, in this outbreak, we improved the identification of asymptomatic carriage with the simple step of additional culture swabs for all cuts and abrasions. Improved identification of colonised recruits, along with an enhanced decolonisation regime and rigid infection control practices, prevented the further spread of the clone through the training centre. In a population with constant ongoing skin trauma, such as the military, contact sport athletes and iIV drug users, our results show that a culture of suitable abrasions/lacerations will improve the identification of MRSA colonisation compared with nasal swabs alone. Despite ongoing skin trauma and the logistical difficulties in delivering effective decolonisation during military training, decolonisation was successful in 79% of recruits after one decolonisation and 87% after the second 10-day decolonisation.

The bacterial pathogen Staphylococcus aureus harbors numerous virulence factors that impact infection severity. Beyond virulence gene presence or absence, the expression level of virulence proteins is known to vary across S. aureus lineages and isolates. However, the impact of expression level on severity is poorly understood due to the lack of high-throughput quantification methods of virulence proteins.
We present a targeted proteomic approach able to monitor 42 staphylococcal proteins in a single experiment. Using this approach, we compared the quantitative virulomes of 136 S. aureus isolates from a nationwide cohort of French patients with severe community-acquired staphylococcal pneumonia, all requiring intensive care. We used multivariable regression models adjusted for patient baseline health (Charlson comorbidity score) to identify the virulence factors whose in vitro expression level predicted pneumonia severity markers, namely leukopenia and hemoptysis, as well as patient survival.
We found that leukopenia was predicted by higher expression of HlgB, Nuc, and Tsst-1 and lower expression of BlaI and HlgC, while hemoptysis was predicted by higher expression of BlaZ and HlgB and lower expression of HlgC. Strikingly, mortality was independently predicted in a dose-dependent fashion by a single phage-encoded virulence factor, the Panton-Valentine leucocidin (PVL), both in logistic (OR 1.28; 95%CI[1.02;1.60]) and survival (HR 1.15; 95%CI[1.02;1.30]) regression models.
These findings demonstrate that the in vitro expression level of virulence factors can be correlated with infection severity using targeted proteomics, a method that may be adapted to other bacterial pathogens.

Many studies have been published assessing the association between the presence of S. aureus genes and outcomes in patients with bone and joint infections (BJI), but it is not known if they have had similar findings. A systematic literature review was performed. All available data on studies in Pubmed between January 2000 to October 2022 reporting the genetic characteristics of S. aureus and the outcomes of BJIs were analyzed. BJI included prosthetic joint infection (PJI), osteomyelitis (OM), diabetic foot infection (DFI), and septic arthritis. Because of the heterogeneity of studies and outcomes, no meta-analysis was performed. With the search strategy, 34 articles were included: 15 articles on children and 19 articles on adults. In children, most BJI studied were OM (n = 13) and septic arthritis (n = 9). Panton Valentine leucocidin (PVL) genes were associated with higher biological inflammatory markers at presentation (n = 4 studies), more febrile days (n = 3), and more complicated/severe infection (n = 4). Other genes were reported anecdotally associated with poor outcomes. In adults, six studies reported outcomes in patients with PJI, 2 with DFI, 3 with OM, and 3 with various BJI. Several genes were associated with a variety of poor outcomes in adults, but studies found contradictory results. Whereas PVL genes were associated with poor outcomes in children, no specific genes were reported similarly in adults. Additional studies with homogenous BJI and larger sample sizes are needed.

Panton Valentine leukocidin (PVL) is a virulence factor which is associated with methicillin sensitive and resistant Staphylococcus aureus (MSSA/MRSA) causing skin and soft tissue infections (SSTI). This study aimed to evaluate a novel lateral flow immunoassay (LFI) for PVL detection in S. aureus cultures and to describe their genotypic characterization.
The study was carried out from January-August 2020 in Dubai, United Arab Emirates. S. aureus isolates associated with SSTI were tested for PVL detection using LFI. DNA microarray-based assays were used for molecular characterization including detection of pvl genes.
One-hundred thirty-five patients with a clinical diagnosis of SSTIs were recruited. Sixty-six patients received antibiotics, mostly beta lactams (n=36) and topical fusidic acid (n=15). One-hundred twenty-nine isolates (MRSA: n=43; MSSA: n=86) were tested by LFI and DNA microarrays. All 76 (58.9%) isolates which were unambiguously negative for the PVL in LFI were negative for pvl genes using the DNA microarray. All the LFI PVL positive isolates (n=53) had pvl genes detected. This translates into 100% each for sensitivity, specificity, positive and negative predictive values for the LFI. The LFI typically takes about 15 min inclusive of a 10 min incubation period. Predominant S. aureus clonal complexes (CC) were CC30 (n=18), CC22 (n=13), CC5 (n=12), CC1 (n=11), CC152 (n=8), CC15 (n=7); CC97 (n=7); CC8 and CC20 (n=6 each). Among MRSA, the proportion of pvl-positives (35/43; 81%) was higher than among MSSA (n/N=18/86; 21%). The fusidic acid resistance gene fusC was detected in 14 MRSA (33%) compared to 8 MSSA (9%). A co-carriage of fusC and pvl genes was present in 7 MRSA and in one MSSA.
LFI shows excellent diagnostic accuracy indices for rapid identification of PVL in MSSA/MRSA in a setting with high prevalence of pvl +ve strains. The high occurrence of pvl and fusC genes in MRSA strains causing SSTI is of concern and needs constant surveillance.

BACKGROUND: Spiders, especially those of the genus Loxoceles such as L. rufescens, endemic in Mediterranean regions, are frequently reported as causes of venom poisoning in humans in the south of France. The most common signs consist of cutaneous necrosis presenting initially as inflammatory cellulitis and progressing towards the emergence of a necrotic centre.
METHODS: We report 4 cases, initially considered as spider bites due to their sudden occurrence and pain. Rigorous clinical examination coupled with collection of samples for laboratory analysis ultimately enabled the diagnosis to be corrected to one of suppurative skin infection caused by Staphylococcusaureus producing the cytotoxin Panton Valentine leucocidin.
CONCLUSIONS: These observations highlight the potential for confusion between spider bites and infections with PVL-producing S. aureus.

Rapid detection of Methicillin Resistant Staphylococcus aureus (MRSA) is an important concern for both treatment and implementation of infection control policies. The present study provides an \'in house\' real-time PCR assay to detect directly nuc, pvl, and mecA genes. The assay is able to perform identification of MRSA, Methicillin-Sensitive S. aureus, Methicillin-Resistant Coagulase Negative Staphylococci and the Panton-Valentine leukocidin virulence gene from rectal and pharyngeal swab samples in a screening context. We found an analytical sensitivity of this current Triplex PCR assay of 514 CFU/mL. Analytical specificity was tested with different Gram-positive and Gram-negative species and yielded no false-positive PCR signal. The sensitivity and specificity of the Triplex Real Time PCR were both 100% for these targets when compared with the culture and conventional methods. This assay is readily adaptable for routine use in a microbiology laboratory, as it will enable the implementation of timely and properly guided therapy and infection control strategies.

Staphylococcus aureus is a pathogen associated a different kind of infection. Molecular markers are useful tools to study microbial epidemiology. Twenty two methicillin-resistant S. aureus (MRSA) and 23 methicillin-susceptible S. aureus (MSSA) were studied by mecA gene, SCCmec cassette, Panton Valentine leucocidin (PVL) and spa polymorphism. The clinical data patients were analyzed. MSSA was prevalent in samples different from skin and soft tissue (SST) and in hospitalized patients, whereas MRSA in SST. SCCmec type IV was predominant, followed by type I. Low presence of PVL was found. In MRSA 11 different types of spa were detected, t019 was the most frequent and associated with outpatient, 17 types were found in MSSA and t189 was prevalent. spa t002 was present in MSSA and MRSA. We found 11 types of spa not reported in our country.

Superficial skin and soft tissue infections (SSTIs) are common among the Indigenous population of the desert regions of Central Australia. However, the overall burden of disease and molecular epidemiology of Staphylococcus aureus complicated SSTIs has yet to be described in this unique population.
Alice Springs Hospital (ASH) admission data was interrogated to establish the population incidence of SSTIs. A prospective observational study was conducted on a subset of S. aureus complicated SSTIs (carbuncles and furuncles requiring surgical intervention) presenting during a one month period to further characterize the clinical and molecular epidemiology. High resolution melting analysis was used for clonal complex discrimination. Real-time polymerase chain reaction identifying the lukF component of the Panton Valentine leucocidin (pvl) gene determined pvl status. Clinical and outcome data was obtained from the ASH medical and Northern Territory shared electronic health records.
SSTIs represented 2.1% of ASH admissions during 2014. 82.6% occurred in Indigenous patients (n = 382) with an estimated incidence of 18.9 per 1, 000 people years compared to the non-Indigenous population of 2.9 per 1000, with an incident rate ratio of 6.6 (95% confidence interval 5.1-8.5). Clinical and molecular analysis was performed on 50 isolates from 47 patients. Community-associated methicillin-resistant S. aureus (CA-MRSA) predominated (57% of isolates). The high burden of SSTIs is partly explained by the prevalence of pvl positive strains of S. aureus (90% isolates) for both CA-MRSA and methicillin-susceptible S. aureus (MSSA). ST93-MRSA and CC121-MSSA were the most prevalent clones. SSTIs due to ST93-MRSA were more likely to require further debridement (p = 0.039), however they also more frequently received inactive antimicrobial therapy (p < 0.001).
ST93-MRSA and CC121-MSSA are the dominant causes of carbuncles and furuncles in Central Australia. Both of these virulent clones harbor pvl but the impact on clinical outcomes remains uncertain. The high prevalence of CA-MRSA supports empiric vancomycin use in this population when antimicrobial therapy is indicated. Prompt surgical intervention remains the cornerstone of treatment.