Panton-Valentine Leukocidin Toxin Gene Research Articles

Molecular epidemiology of methicillin-resistant Staphylococcus aureus isolated from pigs in Japan

Methicillin-resistant Staphylococcus aureus (MRSA) is the leading cause of infection in hospitalized patients and can be prevalent in humans and various animal species. In European countries, MRSA isolates belonging to clonal complex 398 have been detected at high rates in pigs. However, the prevalence of MRSA in pigs and farm environments in Japan remains unclear. MRSA isolates were obtained from pigs in slaughterhouses, diseased pigs on farms, imported breeding pigs, and farm dust. We conducted whole-genome sequencing (WGS) and analyzed the molecular epidemiological relationship between these MRSA isolates using core genome multilocus sequence typing (cgMLST). The prevalence rates of MRSA among pigs in slaughterhouses, diseased pigs on farms, imported breeding pigs, and farm dust were 5.2 %, 3.4 %, 28.8 %, and 0.06 %, respectively. ST 398 isolates that classified as ST398/t034 were isolated from pigs from all sources. The results of cgMLST showed that ST398/t034 isolates originating from domestic pigs clustered into the same cluster as the isolates from imported breeding pigs. However, some clusters only included isolates of domestic pig origin. Most MRSA isolates in this study carried resistance genes for aminoglycosides, β-lactams, macrolides, tetracyclines, and zinc. None of the MRSA isolates in this study harbored Panton-Valentine leukocidin toxin genes. Molecular epidemiological analysis suggested a relationship between isolates from slaughter pigs and imported breeding pigs and the presence of MRSA isolates of domestic origin. However, more data are needed for elucidation of the origin of these MRSA variants in the pig industry in Japan.

Mupirocin and Chlorhexidine Genotypic Resistance Found in Staphylococcus aureus Isolated From Young Infants Below 90 Days Old: A Genetic Basis for Eradication Failure.

To investigate the genetic characteristics associated with eradication failure of Staphylococcus aureus in infants below 90 days old. S. aureus isolated from clinical specimen cultures (blood, surgical tissue, or drainage, pus, etc.) and routine screening cultures in the neonatal intensive care unit (nasal and axillary skin swab) from patients below 90 days old were collected prospectively for 1 year, from August 2017 to July 2018. The isolates underwent typing and screening for genes associated with chlorhexidine (qacA/B), quaternary ammonium (smr), and mupirocin resistance (iles mutation, mupA, mupB), as well as Panton-Valentine leukocidin (PVL) toxin. During the study period, 40 nonduplicate isolates were included for analyses, of which 70.0% were methicillin-resistant S. aureus (MRSA). Mupirocin resistance was found in 25% of the total isolates; 17.4% of the colonizers; and 35.3% of the pathogens (P = 0.196). Chlorhexidine resistance gene was found in 3 MRSA isolates colonized in the nares of preterm infants. All isolates harbored the disinfectant quaternary ammonium compound (QAC) resistance gene. PVL toxin gene was found in 57.5%, and the presence of PVL gene among colonizers and pathogens was similar (69.6% vs. 41.2%, P = 0.072). Mupirocin, chlorhexidine, and QAC-resistant MRSAs harboring the PVL toxin gene were found in the nasal carriages of preterm infants. In this highly vulnerable patient population, one-fourth of the isolates harbored mupirocin-resistant genes, and all were resistant to QAC disinfectants. These strains are associated with persistence in both carriage and environmental reservoirs within the hospitals.

Understanding the Fight Against Resistance: Hospital-Acquired Methicillin-Resistant Staphylococcus Aureus vs. Community-Acquired Methicillin-Resistant Staphylococcus Aureus.

Since the identification of Staphylococcus (S.) aureus, penicillin was exclusively used to combat its disastrous toxic effects. Shortly thereafter, resistant strains arose, which were no longer susceptible to penicillin or methicillin treatments. These strands were later identified as methicillin-resistant Staphylococcus aureus (MRSA). Two particular MRSA strands that are discussed below are the hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) strands and the community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) strands. Despite arising from a single bacterium, S. aureus, each of these two strands possesses quite different resistance and virulence factors. These differences contribute to the type of population in which they affect, their ability to resist traditional treatment approaches, and their overall morbidity and mortality rates. We explore these differences by reviewing several review articles published on various reputable scientific online databases. Findings include sources from studies conducted in the United States, China, Nepal, and Uganda, ranging from 2006 to 2019. These resistance and virulence factors, the Staphylococcal cassette cartridge mecA resistance gene (SCCmec) and the Panton-Valentine Leukocidin toxin gene (PVL), were identified and isolated in each of these studies in order to appreciate similarities and differences in how they impact human beings.

Prevalence and Molecular Genetics of Methicillin-Resistant Staphylococcus aureus Colonization in Nursing Homes in Saudi Arabia.

Objective Methicillin-resistant Staphylococcus aureus (MRSA) is one of the main causative agents of nosocomial infections that has posed a major threat to those with compromised immune systems such as nursing home residents. The aim of this study was to determine the rates of MRSA strains and the types of Staphylococcal Cassette Chromosome mec (SCCmec)in nursing homes in Saudi Arabia. Methods A total of 188 nasal swabs were collected from the residents and nursing staff in two nursing homes in Riyadh, Saudi Arabia. All MRSA isolates were tested for antimicrobial susceptibility and analyzed for mecA and SCCmec typing by multiplex PCR assay. Detection of the Panton–Valentine leukocidin (PVL) gene was also tested in all positive MRSA isolates by multiplex PCR using specific primers. Results Among the 188 collected nasal swabs (105 males and 83 females), MRSA colonization rate was 9.04% (11 (5.85%) females and 6 (5.71%) males). About 47% of MRSA were multidrug resistant (MDR) as acquired resistance to beta-lactam, macrolide, and aminoglycoside antibiotics. However, all the MRSA isolates showed susceptibility to vancomycin, tigecycline, and linezolid. All the MRSA isolates (n = 17) were mecA-positive with the SCCmec IVc (n = 7, 41.18%) as the most common SCCmec type followed by SCCmec V (n = 5, 29.41%) and SCCmec IVa (n = 2, 11.76%). The remaining isolates (n = 3) were nontypeable (17.65%). In addition, the PVL toxin gene was only detected in four of the male samples. Conclusion MRSA nasal colonization is a common incident among nursing home residents. The prevalence of community-associated (CA) MRSA (SCCmec IV and V) was more common than hospital-associated (HA) MRSA in our study samples. It is crucial to investigate such rate of incidence, which is a key tool in preventive medicine and would aid in determining health policy and predict emergent outbreaks.

Screening for Panton-Valentine Leukocidin Toxin Genes in Multi-Drug Resistant Methicillin-Resistant <i>Staphylococcus aureus</i> Nasal Isolates from Healthy School Children in Mariental, Namibia

Panton-Valentine leukocidin (PVL) is one of the toxins responsible for increased virulence of Staphylococcus aureus. In school settings where children are in close contact with each other, S. aureus strains, including those that may produce PVL, can be transmitted and spread in the community. Twenty-two multi-drug resistant MRSA nasal isolates from children enrolled in five schools in the town of Mariental and the multi-drug resistant American Type Culture Collection MRSA reference strain S. aureus ATCC 33591 (PVL-negative control) were used for molecular assays. Plasmid deoxyribonucleic acid (DNA) of isolates was amplified by polymerase chain reaction (PCR) and amplified PCR products were electrophoresed on a 2.5% (w/v) agarose gel containing 12 μl 0.5 μg/ml ethidium bromide and 1× TAE (Tris-acetate-EDTA) buffer at 90 volts for 50 minutes. The developed gel was viewed for the PVL-associated lukS and lukF genes that amplified at 151 bp and 406 bp, respectively. Our results indicated that seven nasal isolates had PVL toxin gene(s). From the seven isolates, three were tested positive for both lukS and lukFgenes, one tested positive for only lukS, and three tested positive for only lukF. Two of the isolates harbouring both lukS and lukF genes shared the same antibiotic resistance pattern and one of them could also produce enterotoxin A. One of the isolates with only lukF gene could produce enterotoxins B and C. These toxin-producing isolates can be expected to be more virulent than non-producers. Children should be educated on the importance of regular handwashing with soap and water to prevent the spread of potentially virulent staphylococci amongst them and the wider community. This work warrants a larger study to be carried out to investigate PVL toxin and its associated infections in Staphylococcus from school children in Namibia.

Multiplex PCR based detection of mecA, mecC and PVL gene in analysis of prevalence, circulation, transmission of MSSA/MRSA strains in a tertiary care hospital

Introduction: Staphylococcus aureus a Gram positive cocci causes major and minor infections in humans. It is the most important pathogen causing Hospital acquired and community-acquired (CA) infections. The commonest being the Methicillin resistant Staphylococcus aureus (MRSA) which is due to the insertion of SCC mec into methicillin-susceptible S. aureus (MSSA) lineages. Though, a number of toxins are involved in the pathogenesis of its infections, Panton Valentine leukocidin (PVL) is considered the common virulence factor for community acquired MRSA. Our study aimed to determine the prevalence of mec A and mecC gene and the presence of PVL toxin genes (lukS-PV and lukF-PV) in S. aureus isolates. Materials and Methods: The study was carried out in Department of Microbiology at Chettinad Hospital & Research Institute, a tertiary care center for 3 months. A total of 109 clinical isolates of S.aureus were subjected for the study (Wards and Outpatient departments). The identification and antimicrobial sensitivity testing were done by standard microbiological techniques. Multiplex PCR was used to detect mecA, mecC and PVL genes. Results: Out of 109 Staphylococcus aureus isolates, 24(22%) were MRSA and 85(78%) were Methicillin sensitive. Among MRSA strains, 13(54.1%) were positive for mecA, 8(33.3%) were positive for PVL, 3 (12.5%) were found positive for mecC. Among the 85(78%) MSSA isolates, 3(3.5%) were positive mecA gene and only 1(1.1%) isolate had mecC gene. The isolates from hospital environment were negative for both mecA and mecC but was positive for PVL genes. Conclusion: Our study showed high prevalence among community acquired MRSA isolates. Keywords: Community acquired staphylococcus aureus, MRSA, mecA, mec C, PVL gene, Multiplex PCR.

The Role of Staphylococcus aureus in Mastitis

Breastfeeding women are at risk of developing mastitis during the lactation period. Staphylococcus aureus has emerged as the community-acquired pathogen responsible for virulence (methicillin resistance and Panton-Valentine leukocidin toxin producing). The aim was to compare the microorganisms responsible for mastitis and breast abscesses during breastfeeding. This observational study was conducted with a sample of women (N = 60) admitted to our hospital between 2016 and 2018. Participants affected by mastitis and breast abscess were studied and cared for by a multidisciplinary working group. A diagnostic breast ultrasound identified the pathology. Twenty-six participants (43.3%) were affected by mastitis and 34 (56.7%) by breast abscess. The most common microorganism identified was Staphylococcus aureus (S. aureus; mastitis, n = 13; abscesses, n = 24). Methicillin resistance was identified in 21 (44.7%) S. aureus strains: 17 (80.9%) cases of abscess and four (19.1%) cases of mastitis. The median number of months of breastfeeding was smaller in the methicillin-resistant S. aureus (MRSA) cases (median = 3, range = 1-20 months) than in the methicillin-sensitive S. aureus (MSSA) cases (median = 6.5, range = 3-21 months). The Panton-Valentine leukocidin toxin gene was detected in 12 (25.5%) cases (MRSA, n = 8, 66.7%; MSSA, n = 4, 33.3%). Hospitalization was required more frequently in MRSA (n = 8, 38%; five Panton-Valentine leukocidin positive) than in MSSA cases (n = 5, 19%; one Panton-Valentine leukocidin positive). Four women out of the eight MRSA cases (50%) that were Panton-Valentine leukocidin positive stopped breastfeeding during mammary pathologies, three (37.5%) participants continued breastfeeding until the follow-up recall, and one case was lost at follow-up. Clinical severity was probably complicated by the presence of the Panton-Valentine leukocidin toxin, which required hospitalization more frequently.

Molecular Characterization of Methicillin Resistant Staphylococcus aureus in West Bank-Palestine.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a public health threat and a major cause of hospital-acquired and community-acquired infections. This study aimed to investigate the genetic diversity of MRSA isolates from 2015 to 2017 and to characterize the major MRSA clones and anti-biogram trends in Palestine.Methodology: Isolates were obtained from 112 patients admitted to different hospitals of West Bank and East Jerusalem, originating from different clinical sources. Antibiotic susceptibility patterns, staphylococcal chromosomal cassette mec (SCCmec) typing, and Staphylococcus aureus protein A (spa) typing were determined. Also, a panel of toxin genes and virulence factors was studied, including: Panton-Valentine Leukocidin (PVL), ACME-arcA, Toxic Shock Syndrome Toxin-1 (TSST-1), and Exfoliative Toxin A (ETA).Results: Of the 112 confirmed MRSA isolates, 100% were resistant to all β-lactam antibiotics. Resistance rates to other non- β-lactam classes were as the following: 18.8% were resistant to trimethoprim-sulfamethoxazole, 23.2% were resistant to gentamicin, 34.8% to clindamycin, 39.3% to ciprofloxacin, and 63.4% to erythromycin. All MRSA isolates were susceptible to vancomycin (100%). Of all isolates, 32 isolates (28.6%) were multidrug- resistant (MDR). The majority of the isolates were identified as SCCmec type IV (86.6%). The molecular typing identified 29 spa types representing 12 MLST-clonal complexes (CC). The most prevalent spa types were: spa type t386 (CC1)/(12.5%), spa type t044 (CC80)/(10.7%), spa type t008 (CC8)/(10.7%), and spa type t223 (CC22)/(9.8%). PVL toxin gene was detected in (29.5%) of all isolates, while ACME-arcA gene was present in 18.8% of all isolates and 23.2% had the TSST-1 gene. The two most common spa types among the TSST-1positive isolates were the spa type t223 (CC22)/(Gaza clone) and the spa type t021 (CC30)/(South West Pacific clone). All isolates with the spa type t991 were ETA positive (5.4%). USA-300 clone (spa type t008, positive for PVL toxin gene and ACME-arcA genes) was found in nine isolates (8.0%).Conclusions: Our results provide insights into the epidemiology of MRSA strains in Palestine. We report a high diversity of MRSA strains among hospitals in Palestine, with frequent SCCmec type IV carriage. The four prominent clones detected were: t386-IV/ CC1, the European clone (t044/CC80), Gaza clone (t223/CC22), and the USA-300 clone (t008/CC8).

Staphylococcus aureus carrying lukS/F Panton-Valentine Leukocidin (PVL) toxin genes in hospitals of Lahore city.

Panton Valentine-Leukocidin (PVL) toxin is secreted by Staphylococcus aureus and is mostly associated with skin and soft tissue infections (SSTI). This study aims to find out the prevalence of lukS/F-PV gene, which encode PVL toxin from strains of SSTI, burn wounds and nasal colonizers of out-patients and to measure the antimicrobial susceptibility of S. aureus isolates. This is an analytical observational cross-section study and was conducted from July 2014 to June 2015 at four tertiary care hospitals and PCSIR Laboratories Complex, Lahore, Pakistan. A total of 376 random clinical swabs were collected from SSTI (n = 179), nasal nares (n = 134) and burn wounds (n = 63) from out-patients' departments (OPD). The specimens were cultured on nutrient and mannitol salt agar (MSA) and the organism was identified by catalase, coagulase, and DNase tests. Antimicrobial susceptibility, methicillin, inducible clindamycin, and high-level mupirocin (HLMR) resistance were determined as per CLSI guidelines. Molecular identification of mecA and lukS/F-PV genes was performed by PCR. We isolated 127 S. aureus, where 41 (32.3%) were MRSA and 86 (67.7%) were MSSA. All MRSA carried mecA gene whereas lukS/F-PV gene was found in 21 MRSA and 31 MSSA strains. Overall, a high antimicrobial resistance was found against MRSA and lukS/F-PV positive MSSA. Inducible clindamycin and high-level mupirocin resistance (HLMR) was 23.6% and 19.5% respectively. A high rate of PVL toxin gene was detected among S. aureus strains and a high prevalence of antimicrobial resistant strains was observed.

Distribution of SCCmec Elements and Presence of Panton-Valentine Leukocidin in Methicillin-Resistant Staphylococcusepidermidis Isolated from Clinical Samples in a University Hospital of Isfahan City, Iran.

Coagulase Negative Staphylococcus (CoNS) is considered as a major pathogen of nosocomial infections among immunosuppressed patients. The aim of this study was to identify the types of Staphylococcal Cassette Chromosome mec (SCCmec) and Panton-Valentine Leukocidin (PVL) gene among clinical Methicillin-Resistant S. epidermidis (MRSE) isolates collected from Isfahan. This cross-sectional study was performed from March 2014 to January 2015 at a tertiary care hospital of Isfahan, Iran. Antimicrobial susceptibility tests of S. epidermidis isolates were performed by the disc diffusion method. All the strains were screened for methicillin resistance based on resistance to cefoxitin (30 μg) disc and presence of mecA gene. Determination of SCCmec typing and PVL toxin gene were performed by PCR method. For categorical variables different groups were compared using the Chi-square test or Fisher exact test. A p-value of <0.05 was considered significant for all statistical tests. The frequency of MRSE was 53.8% according to the presence of mecA gene. The overall resistance rate was high with ciprofloxacin (81.4%). PCR analysis showed that 17% (12/70) of MRSE isolate carried the PVL gene and 43% (30/70) were SCCmec type I; 11.4% (8/70) were type II; and 34.2% (24/70) were type IV, whereas, 11.4% (8/70) of the MRSE isolates could not be typed. SCCmec type I was the major type of SCCmec, which indicates an emergence of this SCCmec type in the studied medical centers. Increased prevalence of SCCmec types in community is cause of an increase in antibiotic resistance among microorganisms.

Application of monoclonal antibodies generated against Panton-Valentine Leukocidin (PVL-S) toxin for specific identification of community acquired methicillin resistance Staphylococcus aureus

Panton-Valentine Leukocidin (PVL) produced by community acquired methicillin Staphylococcus aureus (CA-MRSA) involved in skin and soft-tissue infections and necrotizing pneumonia comprised of two fractions, namely PVL S and PVL F. In the present study, three monoclonal antibodies designated as MAb1, MAb9 and MAb10 were generated against recombinant PVL-S (35kDa) protein of S. aureus. All the three MAbs specifically reacted to confirm PVL-S positive strains of S. aureus recovered from clinical samples in Western blot analysis. Similarly all the three MAbs did not show any binding to other tested 14 different pathogenic bacteria belonging to other genera and species in Western blot analysis. Furthermore, a simple dot-ELISA method was standardized for the identification of PVL-S toxin containing S. aureus strains. Initially in dot-ELISA, Protein A (Spa) of S. aureus posed background noise problems due to the non-specific binding of antibodies resulting in false positive reactions. With the addition of 10mM diethylpyrocarbonate (DEPC) along with 5% milk in PBS in the blocking step prevented this non-specific binding of Spa to mouse anti-PVL monoclonal antibodies in dot-ELISA. Once standardized, this simple dot-ELISA was evaluated with nine PVL positive strains recovered from food, environmental and clinical samples and the results were compared with PCR assay for the presence of PVL toxin genes and also with Western blot analysis. A 100% correlation was found between dot-ELISA, PCR assay and Western blot analysis. Collectively our results suggest the newly developed simple dot-ELISA system can be of immense help in providing, rapid detection of the PVL toxin containing S. aureus strains at a relatively low cost and will be a valuable tool for the reliable identification of CA-MRSA.

The Origins of Staphylococcus aureus Isolated from Blepharitis Based on Panton-Valentine Leukocidin and Antibiotic Susceptibility Testing

Introduction: Blepharitis is a common condition, sometimes associated with Staphylococcus aureus. Panton-Valentine Leukocidin (PVL) toxin is a Staphylococcus aureus virulence factor that can be associated with skin and soft tissue infections. Hospital-acquired Staphylococcus aureus infections are generally multi-resistant to antibiotics and PVL-negative. Community-acquired Staphylococcus aureus infections tend to be broadly susceptible to antibiotics and PVL-positive. Though blepharitis is a common clinical diagnosis, the source of infection is not fully understood. Focus on the origin of disease could aid in better treatment and reduction of anti-infective resistance. The purpose of this study is to determine if Staphylococcus aureus blepharitis is predominantly a community-acquired or hospital-acquired infection based on antibiotic susceptibility and PVL testing. Materials and Methods: Fifty-nine de-identified Staphylococcus aureus isolates collected from patients with blepharitis were tested for antibiotic susceptibility by disk diffusion using multiple antibiotics from several different classes. The isolates were also tested for the presence of Staphylococcus aureus nuclear DNA and PVL toxin gene using PCR. Multi-resistance was defined as resistance to 3 or more classes of antibiotics. Results: Of the 59 isolates with PCR-identified Staphylococcus aureus DNA, 13 (22%) were multi-resistant; 12 (20%) were methicillin-resistant; and, 3(5%) were PVL-positive. Ten (17%) were multi-resistant and PVL-negative, consistent with hospital-acquired infection. None of the isolates were broadly susceptible to antibiotics and PVL-positive, which would be consistent with community-acquired infection. Forty-nine isolates (83%) (p=0.0001) could not be designated to either group. Conclusion: Based on PVL and antibiotic susceptibility testing, our results reject the hypothesis that Staphylococcus aureus blepharitis is a community-acquired infection. Staphylococcus aureus blepharitis appears not to be predominantly community- or hospital-acquired based on these parameters.

Emergence of methicillin resistance and Panton‐Valentine leukocidin positivity in hospital‐ and community‐acquired Staphylococcus aureus infections in Beira, Mozambique

The objective of this study was to investigate the antibiotic resistance patterns, including methicillin resistance, inducible macrolide-lincosamide-streptogramin B (MLSB ) resistance and Panton-Valentine leukocidin (PVL) toxin gene carriage among hospital-acquired Staphylococcus aureus (HA-SA) and community-acquired S. aureus (CA-SA), in Beira, Mozambique. In 2010-2011, two prospective surveillance studies were conducted on post-operative and burn wound infections at the Central Hospital of Beira and on skin and soft tissue abscesses at the São Lucas Health Centre. We cultured pus samples, identified suspected S. aureus isolates and performed antimicrobial susceptibility testing, including detection of MLSB resistance. Real-time polymerase chain reaction was used to detect mecA, Martineau and PVL genes. The prevalence of hospital-acquired methicillin-resistant S. aureus (HA-MRSA) infection among 53 inpatients was 15.1%; the prevalence of community-acquired methicillin-resistant S. aureus (CA-MRSA) infection among 100 outpatients was 1.0%. Inducible MLSB resistance was present in 41.7% and 10.7% of HA-SA and CA-SA isolates, respectively. PVL toxin gene was detected in 81.1% of methicillin-susceptible S. aureus (MSSA) compared with 11.1% of methicillin-resistant S. aureus. Our study shows, for the first time in Mozambique, the emergence of HA-MRSA. The prevalence of CA-MRSA was low, whereas the rate of PVL toxin gene carriage in MSSA was high. The high rate of inducible MLSB resistance indicates the importance of performing routine D-tests. Overall, our results show the need of strengthening laboratory facilities to provide microbiological data for both directed therapy and surveillance.

Detection of Epidemic USA300 Community-Associated Methicillin-Resistant Staphylococcus aureus Strains by Use of a Single Allele-Specific PCR Assay Targeting a Novel Polymorphism of Staphylococcus aureus pbp3

In recent years, the dramatic increase in community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections has become a significant health care challenge. Early detection of CA-MRSA is important because of its increased virulence associated with the arginine catabolic mobile element (ACME), Panton-Valentine leukocidin (PVL), and other toxins that may contribute to disease severity. In particular, the USA300 epidemic clone has emerged and now represents the cause of as much as 98% of CA-MRSA skin and soft tissue infections in the United States. Current diagnostic assays used to identify CA-MRSA strains are based on complex multiplex PCRs targeting the staphylococcal cassette chromosome mec (SCCmec) DNA junction, a multitude of genes, and noncoding DNA fragments or on a number of lengthy sequence-typing methods. Here, two nucleotide polymorphisms, G88A and G2047A, that were found to be in strict linkage disequilibrium in the S. aureus penicillin-binding protein 3 (pbp3) gene were also found to be highly associated with the USA300 clone of CA-MRSA. Clinical isolates that contained this pbp3 allele were also positive for the presence of SCCmec type IV, the ACME, and the PVL toxin gene and matched the t008 or t121 molecular spa types, which are associated specifically with the USA300 CA-MRSA clone. A single allele-specific PCR targeting the G88A polymorphism was developed and was found to be 100% sensitive and specific for the detection of USA300 CA-MRSA and 91.5% sensitive and 100% specific for the detection of all CA-MRSA isolates in this study.

Emergence of methicillin-resistant, vancomycin-intermediate Staphylococcus aureus among patients associated with group A Streptococcal pharyngitis infection in southern India

Beyond Staphylococcus aureus being an etiological agent for several serious clinical complications, the foot prints of S. aureus in pharyngitis infection has also been recently recognized. With due response to the fact, a prospective study was conducted between 2009 and 2010 to describe the molecular epidemiology of S. aureus in throat swabs of pharyngitis patients. A total of 63 methicillin-resistant S. aureus (MRSA) and 102 methicillin-susceptible S. aureus (MSSA) isolates were recovered from 265 throat swabs, representing a community-acquired outpatient population from Tamil Nadu, India. Molecular characterization of MRSA was done by two conventional multiplex PCR assays including Panton-Valentine leukocidin (PVL), mecA and nuc genes, and staphylococcal cassette chromosome mec (SCCmec) typing. Among 165 S. aureus isolates, methicillin resistance was observed in 38.2% (n=63), in which 69.8% (n=44/63) of the MRSA along with 55.9% (n=57/102) of MSSA harbored PVL toxin genes. SCCmec typing showed 50.8% of isolates as SCCmec V (n=32), 44.4% as SCCmec III (n=28), and 1.6% as SCCmec types I, II and IVa (n=1). Multilocus sequence typing performed for 26 selected MRSA isolates resulted in 12 different sequence types (ST), including a novel ST2129/SCCmec III, PVL-positive. Ten MRSA isolates were categorized as ST772 (38.5%)/SCCmec V, PVL-positive, and three isolates as ST368 (11.5%)/SCCmec III, PVL-negative. Though the prominent clones of ST772/SCCmec V were multidrug-susceptible worldwide, they were highly multidrug-resistant in the current study, including four clones intermediate to vancomycin. Totally, 10 (15.9%) out of 63 MRSA isolates were documented as vancomycin-intermediate S. aureus (VISA). Collectively, the present study for the first time portrayed the high prevalence of active MRSA pharyngitis infection and also emphasizes an alarming need for discrimination of pharyngeal-asymptomatic carriers of S. aureus from those with an active S. aureus pharyngitis infection.

Emergence of Hospital- and Community-Associated Panton-Valentine Leukocidin-Positive Methicillin-Resistant Staphylococcus aureus Genotype ST772-MRSA-V in Ireland and Detailed Investigation of an ST772-MRSA-V Cluster in a Neonatal Intensive Care Unit

Sequence type 22 (ST22) methicillin-resistant Staphylococcus aureus (MRSA) harboring staphylococcal cassette chromosome mec (SCCmec) IV (ST22-MRSA-IV) has predominated in Irish hospitals since the late 1990s. Six distinct clones of community-associated MRSA (CA-MRSA) have also been identified in Ireland. A new strain of CA-MRSA, ST772-MRSA-V, has recently emerged and become widespread in India and has spread into hospitals. In the present study, highly similar MRSA isolates were recovered from seven colonized neonates in a neonatal intensive care unit (NICU) in a maternity hospital in Ireland during 2010 and 2011, two colonized NICU staff, one of their colonized children, and a NICU environmental site. The isolates exhibited multiantibiotic resistance, spa type t657, and were assigned to ST772-MRSA-V by DNA microarray profiling. All isolates encoded resistance to macrolides [msr(A) and mpb(BM)] and aminoglycosides (aacA-aphD and aphA3) and harbored the Panton-Valentine leukocidin toxin genes (lukF-PV and lukS-PV), enterotoxin genes (sea, sec, sel, and egc), and one of the immune evasion complex genes (scn). One of the NICU staff colonized by ST772-MRSA-V was identified as the probable index case, based on recent travel to India. Seven additional hospital and CA-ST772-MRSA-V isolates recovered from skin and soft tissue infections in Ireland between 2009 and 2011 exhibiting highly similar phenotypic and genotypic characteristics to the NICU isolates were also identified. The clinical details of four of these patients revealed connections with India through ethnic background or travel. Our study indicates that hospital-acquired and CA-ST772-MRSA-V is currently emerging in Ireland and may have been imported from India on several occasions.

Methicillin-ResistantStaphylococcus aureusin Marine Mammals

Methicillin-Resistant<i>Staphylococcus aureus</i>in Marine Mammals

Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus Bloodstream Isolates in Urban Detroit

To gain a better understanding of epidemiology of resistance in Staphylococcus aureus, we describe the molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream isolates in urban Detroit. Bloodstream isolates from July 2005 to February 2007 were characterized. Two hundred ten bloodstream isolates from 201 patients were evaluated. Patient characteristics were as follows: median age, 54 years; 56% male; and 71% African-American. Seventy-six percent of infections were health care associated, with 55% being community-onset infections and 21% hospital acquired, and 24% were community associated. The most common sources were skin/wound (25%), central venous catheters (24%), unknown source (20%), and endocarditis (9%). Ninety percent and 5% of isolates had a MIC of vancomycin of <or=1.0 mg/liter, using automated dilution testing and E-test, respectively. Six percent of isolates showed heteroresistance to vancomycin, all occurring with isolates having a vancomycin E-test MIC of >or=1.5 mg/liter. Results of pulsed-field gel electrophoresis showed 17 strain types. The predominant strains were USA100 (104 isolates) and USA300 (74 isolates). Forty-nine percent of the isolates had staphylococcal cassette chromosome mec II, and 56% had agr II. All USA300 isolates were positive for the Panton-Valentine leukocidin toxin genes and agr I. Forty-seven percent of USA300 bloodstream infections were health care associated (35% community onset and 12% hospital onset). USA300 strains were more common in injection drug users with skin/wound as the predominant source of infection. Thirty percent of the USA100 strains were closely related to vancomycin-resistant Staphylococcus aureus isolates. The results of this study show that vancomycin MICs using automated dilution testing with Vitek-2 and E-test were highly discordant. Most methicillin-resistant S. aureus strains causing bacteremia are health care associated, commonly have MICs of vancomycin that are high within the susceptible range are not detected by routine automated dilution testing, and have significant diversity of molecular characteristics. USA100 strains that are closely related to vancomycin-resistant S. aureus (VRSA) isolates and USA300 strains are common as causes of both hospital and community-onset infection. Infection control measures should focus not only on prevention of the spread of community strains in the hospital but also prevention of the spread of hospital strains associated with VRSA into the community.

Emergence of Community-Associated Methicillin-ResistantStaphylococcus aureusInfection Among Patients With End-Stage Renal Disease

To evaluate the frequency of infections due to community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains among our patients with end-stage renal disease. Prospective observational clinical and laboratory study of patients in 2005. Molecular features of isolates recovered from these patients were compared with those of isolates recovered in 2000 from patients with end-stage renal disease. A 600-bed urban academic medical center. Thirty-two patients with end-stage renal disease and MRSA infection at the time of hospitalization from 2005 were evaluated. For comparison, laboratory analysis was performed for 17 MRSA isolates recovered from patients with end-stage renal disease in 2000. The patients from 2005 were more likely than the patients from 2000 to have infection with strains that carried the staphylococcal cassette chromosome (SCC) mec type IV complex (50% vs 11.8%; relative risk, 4.25 [95% confidence interval, 1.17-25.98]; P = .012) and the Panton-Valentine leukocidin toxin genes (25% vs 0%; P = .038). Eight patients from 2005 were infected with a strain that is identical to MRSA clone USA300 in terms of molecular type and presence of SCCmec type IV and Panton-Valentine leukocidin genes. Among the patients from 2005, those infected with SCCmec type IV strains (ie, CA-MRSA strains) and those infected with SCCmec type II strains (ie, healthcare-associated MRSA [HA-MRSA] strains) were similar with respect to demographic characteristics, risk factors, and outcomes. We documented an increased proportion of infections with CA-MRSA strains, including clone USA300, among our population of patients undergoing dialysis. Patients infected with CA-MRSA strains and HA-MRSA strains were similar with respect to presenting illness and outcomes.

Methicillin-ResistantS. aureusInfections among Patients in the Emergency Department

Methicillin-resistant Staphylococcus aureus (MRSA) is increasingly recognized in infections among persons in the community without established risk factors for MRSA. We enrolled adult patients with acute, purulent skin and soft-tissue infections presenting to 11 university-affiliated emergency departments during the month of August 2004. Cultures were obtained, and clinical information was collected. Available S. aureus isolates were characterized by antimicrobial-susceptibility testing, pulsed-field gel electrophoresis, and detection of toxin genes. On MRSA isolates, we performed typing of the staphylococcal cassette chromosome mec (SCCmec), the genetic element that carries the mecA gene encoding methicillin resistance. S. aureus was isolated from 320 of 422 patients with skin and soft-tissue infections (76 percent). The prevalence of MRSA was 59 percent overall and ranged from 15 to 74 percent. Pulsed-field type USA300 isolates accounted for 97 percent of MRSA isolates; 74 percent of these were a single strain (USA300-0114). SCCmec type IV and the Panton-Valentine leukocidin toxin gene were detected in 98 percent of MRSA isolates. Other toxin genes were detected rarely. Among the MRSA isolates, 95 percent were susceptible to clindamycin, 6 percent to erythromycin, 60 percent to fluoroquinolones, 100 percent to rifampin and trimethoprim-sulfamethoxazole, and 92 percent to tetracycline. Antibiotic therapy was not concordant with the results of susceptibility testing in 100 of 175 patients with MRSA infection who received antibiotics (57 percent). Among methicillin-susceptible S. aureus isolates, 31 percent were USA300 and 42 percent contained pvl genes. MRSA is the most common identifiable cause of skin and soft-tissue infections among patients presenting to emergency departments in 11 U.S. cities. When antimicrobial therapy is indicated for the treatment of skin and soft-tissue infections, clinicians should consider obtaining cultures and modifying empirical therapy to provide MRSA coverage.