Single-Chain Fragment Variables Targeting Leukocidin ED Can Alleviate the Inflammation of Staphylococcus aureus-Induced Mastitis in Mice.

Staphylococcus aureus is a vital bovine mastitis pathogen causing huge economic losses to the dairy industry worldwide. In our previous studies, leukotoxin ED (LukED) was detected in most S. aureus strains isolated from bovine mastitis. Here, four single-chain fragment variables (scFvs) (ZL8 and ZL42 targeting LukE, ZL22 and ZL23 targeting LukD) were obtained using purified LukE and LukD proteins as the antigens after five rounds of bio-panning. The complementarity-determining region 3 (CDR3) of the VH domain of these scFvs exhibited significant diversities. In vitro, the scFvs significantly decreased LukED-induced cell killing by inhibiting the binding of LukED to chemokine receptors (CCR5 and CXCR2) and reduced the death rates of bovine neutrophils and MAC-T cells caused by LukED and S. aureus (p < 0.05). In an S. aureus-induced mouse mastitis model, histopathology and MPO results revealed that scFvs ameliorated the histopathological damages and reduced the infiltration of inflammatory cells (p < 0.05). The ELISA and qPCR assays showed that scFvs reduced the transcription and expression levels of Tumor Necrosis Factor-alpha (TNF-α), interleukin-1ß (IL-1ß), IL-6, IL-8 and IL-18 (p < 0.05). The overall results demonstrated the protective anti-inflammatory effect of scFvs in vitro and in vivo, enlightening the potential role of scFvs in the prevention and treatment of S. aureus-induced mastitis.

Characterisation of clinical Staphylococcus aureus isolates harbouring mecA or Panton-Valentine leukocidin genes from four tertiary care hospitals in Indonesia.

OBJECTIVES: To determine the prevalence, antimicrobial susceptibility profiles and clonal distribution of either methicillin-resistant Staphylococcus aureus (MRSA) or Panton-Valentine leukocidin (PVL)-positive S. aureus obtained from clinical cultures in Indonesian hospitals. METHODS: S. aureus isolates from clinical cultures of patients in four tertiary care hospitals in Denpasar, Malang, Padang and Semarang were included. We assessed the antimicrobial susceptibility profiles using the Vitek2(®) system, determined the presence of the mecA gene and genes encoding PVL using PCR and analysed the clonal relatedness with Raman spectroscopy. SCCmec typing was performed for all MRSA isolates. Multilocus sequence typing (MLST) was performed for a subset of isolates. RESULTS: In total, 259 S. aureus strains were collected. Of these, 17/259 (6.6%) and 48/259 (18.5%) were MRSA and PVL-positive methicillin-susceptible S. aureus (MSSA), respectively. The prevalence of MRSA and PVL-positive MSSA ranged between 2.5-8.9% and 9.5-29.1%, respectively and depended on geographic origin. PVL-positive MRSA were not detected. Raman spectroscopy of the strains revealed multiple Raman types with two predominant clusters. We also showed possible transmission of a ST239-MRSA-SCCmec type III strain and a ST121 PVL-positive MSSA in one of the hospitals. CONCLUSIONS: We showed that MRSA and PVL-positive MSSA are of clinical importance in Indonesian hospitals. A national surveillance system should be set-up to further monitor this. To reduce the prevalence of MRSA in Indonesian hospitals, a bundle of intervention measures is highly recommended.

Staphylococcus aureus leukotoxin GH promotes formation of neutrophil extracellular traps.

Staphylococcus aureus secretes numerous virulence factors that facilitate evasion of the host immune system. Among these molecules are pore-forming cytolytic toxins, including Panton-Valentine leukocidin (PVL), leukotoxin GH (LukGH; also known as LukAB), leukotoxin DE, and γ-hemolysin. PVL and LukGH have potent cytolytic activity in vitro, and both toxins are proinflammatory in vivo. Although progress has been made toward elucidating the role of these toxins in S. aureus virulence, our understanding of the mechanisms that underlie the proinflammatory capacity of these toxins, as well as the associated host response toward them, is incomplete. To address this deficiency in knowledge, we assessed the ability of LukGH to prime human PMNs for enhanced bactericidal activity and further investigated the impact of the toxin on neutrophil function. We found that, unlike PVL, LukGH did not prime human neutrophils for increased production of reactive oxygen species nor did it enhance binding and/or uptake of S. aureus. Unexpectedly, LukGH promoted the release of neutrophil extracellular traps (NETs), which, in turn, ensnared but did not kill S. aureus. Furthermore, we found that electropermeabilization of human neutrophils, used as a separate means to create pores in the neutrophil plasma membrane, similarly induced formation of NETs, a finding consistent with the notion that NETs can form during nonspecific cytolysis. We propose that the ability of LukGH to promote formation of NETs contributes to the inflammatory response and host defense against S. aureus infection.

[Laboratory-based evaluation of PVL-RPLA "Seiken" to detect Panton-Valentine leukocidin produced by Staphylococcus aureus].

It is well known that some isolates of Staphylococcus aureus produce pathogenic toxin, Panton-Valentine leukocidin (PVL), and that the toxin has been reported to be highly associated with community acquired methicillin resistant S. aureus (CA-MRSA). Currently, the PCR method using specific primers for the PVL gene (LukS-PV-lukF-PV) have been widely used to detect PVL. In this study, we evaluated the PVL-RPLA "Seiken", diagnostic reagent based on a reserved passive latex agglutination reaction with a specific monoclonal antibody for detecting PVL. A total of 630 clinical isolates were used. PCR method detected 34 PVL-positive (28 MRSA and 6 MSSA), and, of these, PVL-RPLA "Seiken" read positive for 32 isolates (27 MRSA and 5 MSSA), the result indicating two false negative occurrences. The concordance rate was 99.7%. In addition the recommended BHI broth, CCY medium, Dolman broth and Todd-Hewitt broth were applied for toxin preparation media. Toxin concentration produced in CCY medium was significantly higher than those in the remaining culture medium (p < 0.05). PVL-RPLA "Seiken" is a method for detecting the PVL in the culture broth by antigen antibody reaction after an overnight shaking culture. This method does not require any expensive equipments or facilities. Thus this reagent provides us with rapid, easy-to-perform, less expensive test method to detect PVL in clinical microbiology laboratories.

Isolation of methicillin-resistant Staphylococcus aureus (MRSA) from retail meats in Hong Kong.

OBJECTIVES: The presence of methicillin-resistant Staphylococcus aureus (MRSA) on meat purchased from retail outlets may allow its spread to households and represents a risk for colonization and possibly infection of consumers. Improved isolation methods have indicated that more than 10% of samples are positive. We aimed to determine rates of MRSA contamination of meat samples, including comparison of fresh and frozen samples. We characterized isolates and determined their antibiotic susceptibility. METHODS: Samples of raw meats commonly consumed in Hong Kong were investigated for MRSA contamination using a double-enrichment isolation method. Isolates were characterized by antibiotic susceptibility testing, presence of mecA, SCCmec type, staphylococcal enterotoxins, Panton-Valentin leukocidin (PVL), and spa type. Differences in rates of MRSA contamination between meat types, rearing method, locations, sources, and fresh or frozen storage were compared. RESULTS: MRSA was recovered from 21.9% of pork samples (78/355), 6.8% chicken (31/455), and 4.4% of beef (17/380). Isolation was considerably higher from fresh pork (47%) than frozen (0.6%), whereas contamination rates in fresh (6%) and frozen (7%) chicken were similar. All strains were multidrug resistant. All contaminated fresh pork and most frozen chicken originated from China. Most isolates belonged to CC9, being SCCmec IVb and spa type t899 or closely related spa types, but one chicken sample yielded ST398. Five strains carried spa types associated with human isolates. The egc enterotoxin group was present in the majority of isolates, but PVL in only three from chicken. CONCLUSIONS: The predominance of t899 in isolates indicates that the primary source of contamination may be pig carcasses, previously demonstrated to frequently harbor CC9-positive MRSA in Hong Kong and China. The high rates of meat contamination suggest that improvements in food safety and personal hygiene guidelines may be advisable to reduce risk of spread of these MRSA strains in the community.

Panton-Valentine leukocidin positive sequence type 80 methicillin-resistant Staphylococcus aureus carrying a staphylococcal cassette chromosome mec type IVc is dominant in neonates and children in an Algiers hospital.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major antimicrobial drug-resistant pathogen causing serious infections. It was first detected in healthcare settings, but in recent years it has also become disseminated in the community. Children and young adults are most susceptible to infection by community-acquired (CA) MRSA strains. In this study 25 MRSA isolates implicated in infections of neonates and children admitted to an Algiers hospital during an 18 month period were characterized by molecular methods including staphylococcal cassette chromosome (SCC) mec typing, PCR amplification of pvl genes, pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Fifteen out of 25 isolates were from hospital-acquired infections. Twenty-four isolates carried SCCmec type IVc and belonged to the sequence type (ST) 80, one isolate carried SCCmec type II and was ST 39. Twenty-two out of 24 ST80-MRSA-IVc isolates carried pvl genes. Our results suggest that the Panton-Valentine leukocidin positive ST80- MRSA-IVc is the dominant MRSA clone causing disease in neonates and children in Algiers.

Frequency and clinical association of Panton-Valentine leukocidin-positive Staphylococcus aureus isolates: a study from Kuwait.

OBJECTIVE: This study was undertaken to determine the frequency of Panton-Valentine leukocidin (PVL)-producing Staphylococcus aureus among strains isolated in our laboratory and to study the association of PVL-positive strains with clinical disease. MATERIALS AND METHODS: A total of 291 S. aureus isolates obtained from different clinical specimens from June 1, 2009, to March 31, 2010, at the Farwania Hospital Laboratory were investigated for antimicrobial susceptibility, carriage of genes for PVL, and SCCmec elements. Antimicrobial susceptibility testing was performed by standard methods. The presence of mecA genes for PVL SCCmec typing was determined by PCR. RESULTS: Of the 291 S. aureus isolates, 89 (30.6%) were methicillin-resistant S. aureus (MRSA), whereas 202 (69.4%) were methicillin susceptible (MSSA). Genes for PVL were detected in 13 (14.6%) and 24 (12.0%) of the MRSA and MSSA isolates, respectively. The majority of the PVL-producing MRSA and MSSA were isolated from 12 (30.7%) and 19 (21.8%) cases of skin and soft tissue infections (SSTI), respectively. Although both MSSA and MRSA strains were uniformly susceptible to rifampicin, teicoplanin, and vancomycin, multidrug resistance was observed among PVL-producing and nonproducing MRSA isolates. Both MRSA types carried SCCmec type III, IV, IVc, and V genetic elements. CONCLUSION: This study revealed the presence of genes for PVL in both MSSA and MRSA, associated mostly with SSTI and respiratory tract infections, supporting previous observations that PVL production is widespread among S. aureus strains obtained from different clinical sources.

Drug-resistant human Staphylococcus aureus in sanctuary apes pose a threat to endangered wild ape populations.

Reintroduction of sanctuary apes to natural habitat is considered an important tool for conservation; however, reintroduction has the potential to endanger resident wild apes through the introduction of human pathogens. We found a high prevalence of drug-resistant, human-associated lineages of Staphylococcus aureus in sanctuary chimpanzees (Pan troglodytes) from Zambia and Uganda. This pathogen is associated with skin and soft tissue diseases and severe invasive infections (i.e. pneumonia and septicemia). Colonization by this bacterium is difficult to clear due to frequent recolonization. In addition to its pathogenic potential, human-related S. aureus can serve as an indicator organism for the transmission of other potential pathogens like pneumococci or mycobacteria. Plans to reintroduce sanctuary apes should be reevaluated in light of the high risk of introducing human-adapted S. aureus into wild ape populations where treatment is impossible.

Epidemiology of Staphylococcus aureus in pigs and farmers in the largest farm in Dakar, Senegal.

Between December 2009 and November 2011, we collected 57 (12.3%) Staphylococcus aureus isolates from 464 pigs and 16 (30.8%) isolates from 52 farmers in the largest farm in Dakar. Fifty-one isolates (70%) belonged to four major multilocus sequence typing clonal complexes (CCs): CC152 (26.0%), CC15 (19.2%), CC5 (13.7%), and CC97 (10.9%). The CC variability among the pigs was similar to that observed among the farmers. Six isolates that were recovered only among pigs were resistant to methicillin (10.5%). They were assigned to the ST5-staphylococcal cassette chromosome mec type (SCCmec) IV (n = 5) and ST88-SCCmec IV (n = 1) clones. The luk-PV genes encoding Panton-Valentine leukocidin (PVL), present in 43 (58.9%) isolates overall, including all major CCs and the MRSA ST5-SCCmec IV clone, were highly prevalent compared to data from industrialized countries. This finding is of major concern with regard to the potential virulence of these strains.

Cases of community-acquired meticillin-resistant Staphylococcus aureus in an asylum seekers centre in Germany, November 2010.

In an asylum seeker centre in Schleswig-Holstein, a resident was diagnosed with furuncle caused by a Panton-Valentine leukocidine (PVL)-positive community-acquired meticillin-resistant Staphylococcus aureus (CA-MRSA). As a result of active case finding, 232 of 427 persons (54% of all residents) were screened for MRSA and two further PVL-positive CA-MRSA cases were identified

Skin and soft tissue infections caused by community-associated methicillin-resistant staphylococcus aureus among children in China.

AIM: To investigate the characteristic of community-associated methicillin-resistant staphylococcus aureus (CA-MRSA) skin and soft tissue infections (SSTIs) among children in China. METHODS: Forty-seven children with CA-MRSA SSTIs were enrolled in this study. Clinical information was collected and analysed. The strains from the children were analysed by multilocus sequence typing (MLST), staphylococcus cassette chromosome mec (SCCmec) typing and spa typing. The Panton-Valentine leukocidin (PVL) gene was also detected. RESULTS: The majority of the 47 cases were impetigo (20; 42.6%) and abscesses (14; 29.8%). The rest was cellulites, infected wounds, omphalitis, paronychia and conjunctivitis combined folliculitis. Thirty-two of the isolates (68.1%) were PVL-positive, and the abscesses infected with PVL-positive strains usually required incision and drainage (87.5% vs. 16.7%, p = 0.026). Most of the isolates belonged to ST type 59, which accounted for 46.8%, followed by ST1 (7/47, 14.9%) and ST910 (5/47, 10.6%). The clone of ST59-MRSA-IV with t437 was the most prevalent one. The multiresistant rate of these strains was 93.6%. CONCLUSION: The most common disease of CA-MRSA SSTIs was impetigo, and PVL-positive abscess was associated with incision and drainage. ST59-MRSA-IV with t437 was the most prevalent clone, and the multiresistant rate was high in Chinese children.

A pentaplex PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus and Panton-Valentine Leucocidin.

BACKGROUND: Staphylococcus aureus is a major human pathogen, especially methicillin-resistant S. aureus (MRSA), which causes a wide range of hospital and community-acquired infections worldwide. Conventional testing for detection of MRSA takes 2-5 days to yield complete information of the organism and its antibiotic sensitivity pattern. RESULTS: The present study focused on the development of a pentaplex PCR assay for the rapid detection of MRSA. The assay simultaneously detected five genes, namely 16S rRNA of the Staphylococcus genus, femA of S. aureus, mecA that encodes methicillin resistance, lukS that encodes production of Panton-Valentine leukocidin (PVL), a necrotizing cytotoxin, and one internal control. Specific primer pairs were successfully designed and simultaneously amplified the targeted genes. The analytical sensitivity and specificity of the pentaplex PCR assay was evaluated by comparing it with the conventional method. The analytical sensitivity of the pentaplex PCR at the DNA level was found to be 10 ng DNA. The analytical specificity was evaluated with 34 reference staphylococci and non-staphylococcal strains and was found to be 100%. The diagnostic evaluation of MRSA carried out using 230 clinical isolates, showed 97.6% of sensitivity, 99.3% of specificity, 98.8% of positive predictive value and 98.6% of negative predictive value compared to the conventional method. The presence of an internal control in the pentaplex PCR assay is important to exclude false-negative cases. CONCLUSION: The pentaplex PCR assay developed was rapid and gave results within 4 h, which is essential for the identification of Staphylococcus spp., virulence and their resistance to methicillin. Our PCR assay may be used as an effective surveillance tool to survey the prevalence of MRSA and PVL-producing strains in hospitals and the community.

Pleuropulmonary complications of PVL-positive Staphylococcus aureus infection in children.

UNLABELLED: It is increasingly recognized world-wide that Panton-Valentine leukocidin (PVL)-positive Staphylococcus aureus (PVL-SA) is associated with a highly aggressive and often fatal form of community-acquired pneumonia. We report four children who presented with severe pleuropulmonary complications due to infection by community-acquired methicillin-sensitive S. aureus (CA-MSSA), producing PVL toxin. The complications included bilateral multilobular infiltrates, pneumatocoeles, recurrent pneumothoraces, pleural effusion, empyema, lung abscess and diaphragmatic paralysis. This case series highlights the diverse pleuropulmonary manifestations of this potentially lethal infection and the importance of heightened awareness, early recognition and aggressive therapy. CONCLUSION: Complicated pneumonia in a previously fit young patient with a history of preceding 'flu-like' illness or skin/soft tissue infection should raise the suspicion of infection by PVL-positive Staphylococcus aureus (PVL-SA). Severe pleuropulmonary complications are a feature of this disease.

Prevalence and distribution of meticillin-resistant Staphylococcus aureus within the environment and staff of a university veterinary clinic.

OBJECTIVES: To characterise the distribution of meticillin-resistant Staphylococcus aureus within the environment of a university small animal hospital and compare this with the distribution among staff. METHODS: Samples were collected from 140 environmental sites and the anterior nares of 64 staff members at the University of Glasgow Small Animal Hospital on a single day (d1). Sixty of the environmental sites were resampled 14 days later (d14). RESULTS: Meticillin-resistant S aureus was isolated from two of 140 (1.4 per cent; 95 per cent confidence interval: 1.7 to 5.1) environmental sites on d1 and one of 60 (1.7 per cent; 95 per cent confidence interval: 0.4 to 8.9) on d14. Two of the 64 staff sampled were positive for meticillin-resistant S aureus (3.1 per cent; 95 per cent confidence interval: 0.4 to 8.4). CLINICAL SIGNIFICANCE: A lower prevalence of meticillin-resistant S aureus was observed in the environment than previously reported. The location, relatedness between isolates and the presence of Panton-Valentine leucocidin indicates that the source of the environmental meticillin-resistant S aureus was most likely to have been human rather than animal in these cases. This study presents important information regarding the potential source and distribution of meticillin-resistant S aureus within veterinary hospital environments and highlights potential variability of prevalence of meticillin-resistant S aureus within and between veterinary institutions.

PVL Staphylococcus aureus osteomyelitis complicating septic arthritis in a UK soldier serving in Iraq.

Musculoskeletal infections caused by Panton-Valentine Leukocidin (PVL) secreting Stapylococcus aureus in children and adolescents have previously been reported. We report the first adult case in a 26 year-old British Army soldier who presented with a S. aureus septic arthritis. He was treated by surgical washout and antibiotics and discharged but was readmitted five months later with an ipsilateral femoral osteomyelitis requiring debridement. The causative S. aureus grown from tissue biopsy taken at time of surgery was found to encode the PVL gene. Whilst there is evidence that soldiers in Iraq have a greater rate of S. aureus colonisation on their skin, the proportion that encode the PVL gene is similar to that observed in the UK. Soldiers are however, subject to the known risk factors that increase vulnerability to PVL secreting S. aureus infection. Military clinicians need to be aware of PVL secreting S. aureus and have a low threshold for requesting specific testing in aggressive musculoskeletal S. aureus infections.

Methicillin-resistant Staphylococcus aureus nasal carriage in international medical conference attendees.

BACKGROUND: Carriage of methicillin-resistant Staphylococcus aureus (MRSA) is associated with its transmission. International travels and massive gatherings may accelerate such transmission. MRSA carriage was surveyed among the attendees of two international medical conferences held in Taipei in 2010. METHODS: A total of 209 attendees from 23 countries were recruited. Nasal specimens were collected from each volunteer and subjected to polymerase chain reaction (PCR) detection for MRSA. Molecular analysis, including pulsed-field gel electrophoresis, multilocus sequence typing (MLST), typing of staphylococcal cassette chromosome mec (SCCmec) and staphylococcal protein A (spa) genes, and detection of Panton-Valentine leukocidin (PVL) and sasX genes, was performed. RESULTS: MRSA carriage was detected in 10 (4.8%) attendees from Vietnam (3/8, 37.5%), Korea (2/6, 33.3%), Japan (2/41, 4.9%), Philippines (2/52, 3.8%), and Bangladesh (1/4, 25.0%). The proportion of MRSA colonizers was significantly higher in the local hospital group compared to those from the other groups (3/17 vs. 7/192, p < 0.05). Six MRSA isolates were available for molecular analysis. They all carried a type IV SCCmec gene. Five pulsotypes were identified; four genotypes, respectively, were identified by MLST and spa typing. None of the isolates carried either PVL or sasX genes. None of common molecular characteristics was shared by isolates from different countries. Most of these isolates were local endemic community clone in each country. CONCLUSIONS: As healthcare workers, a certain proportion of international medical conference attendees harbored MRSA in their nares, mostly local endemic community clones in each country, which has the potential of spread among attendees.

Genetic characterization of Staphylococcus aureus isolates carrying Panton-Valentine leukocidin genes in Bangladesh.

To assess the spread and genetic characteristics of Panton-Valentine leukocidin (PVL) gene-carrying Staphylococcus aureus in Bangladesh, we investigated 59 strains (49 isolates from clinical specimens and 10 isolates colonized in the nasal cavities of medical staff), including 26 methicillin-resistant S. aureus (MRSA) strains. The PVL gene was detected only in methicillin-susceptible S. aureus (MSSA) strains (7 clinical strains and 2 colonizing strains). PVL gene-positive MSSA strains were found to belong to coagulase serotypes III or VI and were classified into sequence types ST88 (CC88), ST772, and ST573 (CC1) by multilocus sequence typing, and agr types 2 or 3. These types were different from those determined for MRSA (coagulase serotypes I and IV, ST240 and ST361, and agr type 1). PVL gene-positive MSSA possessed a larger number of virulence factor genes than MRSA, although they were susceptible to more antimicrobials. These findings suggest that the PVL gene is distributed to limited populations of S. aureus clones with specific genetic traits that are distinct from MRSA in Bangladesh, but genetically close to CA-MRSA clones in the CC1 lineage reported in the United States and European countries.

Serious life-threatening multifocal infection in a child, caused by Panton-Valentine leucocidin-producing Staphylococcus aureus (PVL-MSSA).

Groin pain is a frequently occurring complaint in presentations to the Emergency Department. Muscular sprain is often a differential diagnosis, however serious conditions such as pyomyositis should not be ignored. This case report presents a child with atraumatic right groin pain, which was initially diagnosed as a muscular sprain. The patient later re-presented out of hours to the Emergency Department with what was found to be extensive pelvic abscesses. He was subsequently found to have bilateral pneumonia and later developed a pericardial effusion and osteomyelitis of the right iliac bone, sacroiliac joint and sacrum. With multiple surgical interventions and appropriate antibiotics, he made a full recovery and was discharged home after a total admission time of 41 days. The causative organism was found to be Panton-Valentine leucocidin-positive methicillin-susceptible Staphylococcus aureus.

Virulence Factors in Staphylococci Isolated From Nasal Cavities of Footballers.

AIM: This study aimed to investigate the rate of Panton-Valentine Leukocidin producing Staphylococcus aureus and methicillin (mecA) and slime (icaA/icaD) genes in staphylococcal strains isolated from nasal cavities of footballers. MATERIALS AND METHODS: Nasal swab samples were taken from each footballers and a healthy control group for the isolation of staphylococcal strains. The polymerase chain reaction technique was used to determine Panton-Valentine Leukocidin, mecA and icaA/icaD genes in staphylococcal isolates. RESULTS: Among 91 S. aureus strains, the presence of mecA gene was detected as 9.9%. This ratio was 17.9% (27 of 151) among the coagulase-negative staphylococci. A significant difference was found between coagulase-negative staphylococci and S. aureus isolates regarding the presence of mecA gene (P < 0.001). As for the genes of the slime, icaA/icaD genes were detected in 198 of 242 (81.8%) strains. The occurrence of slime genes was 91.2% and 89.4% among the S. aureus coagulase and negative staphylococci, respectively (P > 0.05). There was a statistically significant difference between the frequency of the mecA and slime genes when compared with the healthy control group and the football players (P < 0.01). Of 91 isolates, 22 were found to be methicillin resistant by the oxacillin disc diffusion method, whereas the remaining (220) were methicillin susceptible. Methicillin resistance was detected as 14.9% by the polymerase chain reaction method, whereas it was found as 9.1% by phenotypic methods. CONCLUSIONS: Early and accurate diagnosis of virulent staphylococcal strains is crucial because the virulent coagulase-negative and coagulase-positive staphylococcal strains in the nasal floras of footballers may be major potential sources of superficial and deep tissue infections.

Enterovirus D68 and Panton-Valentine Leukocidin-Positive Staphylococcus aureus Respiratory Coinfection with Fatal Outcome.

A previously healthy 10-year-old girl with a 2-day history of upper respiratory illness and fever rapidly developed respiratory failure and sepsis with leukopenia, and expired despite attempts at resuscitation. Postmortem examination revealed bilateral necrotizing pneumonia and evidence of disseminated intravascular coagulation. Nasopharyngeal swabs and lung tissue submitted to the Centers for Disease Control and Prevention (CDC) were positive for Enterovirus D68 (EV-D68). Blood and lung cultures were positive for methicillin-resistant Staphylococcus aureus (MRSA). The isolates were submitted to the CDC and were found to be positive for the toxin Panton-Valentine leukocidin. We describe a fatality related to invasive toxin-mediated MRSA associated with EV-D68 coinfection, along with the clinical, laboratory, and autopsy findings, which provided important clues, prompting further investigation at the CDC to arrive at the correct diagnosis.