Presence Of Pvl Genes Research Articles

Atopic dermatitis pediatric patients show high rates of nasal and intestinal colonization by methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci

BackgroundAtopic dermatitis (AD) patients have high rates of colonization by Staphylococcus aureus, which has been associated with worsening of the disease. This study characterized Staphylococcus spp isolates recovered from nares and feces of pediatric patients with AD in relation to antimicrobial susceptibility, staphylococcal cassette chromosome mec (SCCmec) type, presence of pvl genes and clonality. Besides, gut bacterial community profiles were compared with those of children without AD.ResultsAll 55 AD patients evaluated had colonization by Staphylococcus spp. Fifty-three (96.4%) patients had colonization in both clinical sites, whereas one patient each was not colonize in the nares or gut. Staphylococcus aureus was identified in the nostrils and feces of 45 (81.8%) and 39 (70.9%) patients, respectively. Methicillin-resistant Staphylococcus spp. isolates were found in 70.9% of the patients, and 24 (43.6%) had methicillin-resistant S. aureus (MRSA). S. aureus (55.6%) and S. epidermidis (26.5%) were the major species found. The prevalent lineages of S. aureus were USA800/SCCmecIV (47.6%) and USA1100/SCCmecIV (21.4%), and 61.9% of the evaluated patients had the same genotype in both sites. Additionally, gut bacterial profile of AD patients exhibits greater dissimilarity from the control group than it does among varying severities of AD.ConclusionsHigh rates of nasal and intestinal colonization by S. aureus and methicillin-resistant staphylococci isolates were found in AD patients. Besides, gut bacterial profiles of AD patients were distinctly different from those of the control group, emphasizing the importance of monitoring S. aureus colonization and gut microbiome composition in AD patients.

Antimicrobial Resistance and Virulence Gene Profile of Clinical Staphylococcus aureus: A Multi-Center Study from Ethiopia.

Staphylococcus aureus causes a wide range of infections from mild skin and soft tissue to severe life-threatening bacteremia. The pathogenicity of S. aureus infections is related to various bacterial surface components and extracellular proteins such as toxic-shock syndrome (TSS) toxin and Panton-Valentine leukocidin (PVL). In this study we determine the antimicrobial resistance of isolated strains and their virulence genes in Ethiopia. A total of 190 archived S. aureus isolates from four Ethiopia Antimicrobial Resistance (AMR) Surveillance sites were analyzed. The identification of S. aureus was done by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF Biotyper) and antimicrobial susceptibility test (AST) was done using VITEK® 2. Multiplex PCR was used to detect mecA, mecC, pvl and spa genes and super-antigens (sea, seb, sec, seh and sej staphylococcal enterotoxins). A total of 172 isolates were confirmed as S. aureus, 9 (5.23%) were methicillin-resistant S. aureus (MRSA) and 163 (94.76%) were methicillin-susceptible S. aureus (MSSA). AST showed that 152 (88.4%) isolates were resistant to penicillin; 90 (52.32%) resistant to trimethoprim-sulfamethoxazole; and 45 (26.16%) resistant to tetracycline. A total of 66 (38.37%) isolates harbored at least one staphylococcal enterotoxin gene and 31 (46.96%) isolates had more than one. The most frequent enterotoxin gene encountered was seb 28 (16.28%). The TSST-1 gene was detected in 23 (13.37%). Presence of staphylococcal enterotoxin gene showed significant association with antibiotic resistance to cefoxitin, benzylpenicillin, oxacillin, erythromycin, clindamycin, tetracycline and SXT. The pvl gene was detected in 102 (59.3%) of isolates. Isolates from patients below 15 years of age showed significantly high numbers of pvl gene (P = 0.02). Presence of sej (P = 0.011) and TSST-1 (P <0.001) genes were associated with the presence of pvl gene. In this study, isolates were highly resistant to oral antibiotics and the pvl, seb, sea and TSST-1 genes were prevalent.

Study on PVL, blaOXA-23 and blaOXA-51 Genes in Drug Resistant &amp;lt;i&amp;gt;Staphylococcus aureus&amp;lt;/i&amp;gt; Causing Surgical-Sites and Traumatic Wounds Infections, Sudan

Background: The characteristics of Staphylococcus aureus that made it the most important cause of wound infections are environmental spread antimicrobials resistance and virulence. Absence of molecular detection of drug resistance and virulence factors in many developing countries limits the epidemiological information. This study conducted to identify PVL virulence gene, and blaOXA-23 and blaOXA-51 drug resistance genes of Staphylococcus aureus isolated from surgical-sites infections (SSIs) and traumatic wounds. Methods: A cross-sectional study was conducted from 2019 to 2021, in which 70 cefepime resistant Staphylococcus aureus were used, the strains were isolated from patients of SSIs and traumatic wounds admitted to the department of General Surgery in Wad Medani Teaching Hospital. Mannitol salt agar was used for primary culture followed by biochemical identification and Kirby Bauer susceptibility testing. Single and multiplex PCR protocols performed for bacterial confirmation and target genes detection. Results: Staphylococcus aureus strains from SSIs constituted 56% (39/70) from which 41% (16/39) possessed PVL gene while 42% (13/31) of wound infections strains were positive for PVL gene. Presence of PVL gene was significantly associated with resistance to meropenem (P. value 0.023) and ceftriaxone (P. value 0.037). blaOXA-23 was significantly detected with resistance to meropenem, augmentin and ceftriaxone. While blaOXA-51 was significantly identified among Staphylococcus aureus strains that showed resistance to meropenem and ciprofloxacin. Conclusion: This is the first study in Sudan that identified blaOXA-23 and blaOXA-51 in Staphylococcus aureus and correlated them to resistance to commonly used antimicrobials. Meropenem resistant Staphylococcus aureus were significantly positive for PVL, blaOXA-23 and baOXA-51 genes.

Differences in gram-positive bacterial colonization and antimicrobial resistance among children in a high income inequality setting

BackgroundStaphylococcus aureus and beta-hemolytic streptococci (BHS) diseases disproportionately affect populations in middle/low-income countries. To assess if this disparity is reflected in colonization by these organisms, we compared their colonization frequency among children from different socioeconomic status (SES) communities in a city with high income inequality.MethodsBetween May–August 2014, we collected nasal and throat swabs to investigate S. aureus and BHS colonization among children who attended private and public pediatric clinics. Patients were classified as high SES, middle/low SES, and slum residents. We investigated the antimicrobial resistance profile, the SCCmec types and the presence of PVL genes among methicillin-resistant S. aureus (MRSA). We also examined the antimicrobial resistance profile and serogroups of BHS.ResultsOf 598 children, 221 (37%) were colonized with S. aureus, of which 49 (22%) were MRSA. MRSA colonization was higher in middle/low SES (n = 18; 14%) compared with high SES (n = 17; 6%) and slum (n = 14; 8%) residents (p = 0.01). All MRSA strains were susceptible to clindamycin, nitrofurantoin, and rifampin. The highest non-susceptibility frequency (42.9%) was observed to erythromycin. SCCmec type V was only found in isolates from high SES children; types I and II were found only in middle/low SES children. Ten (20%) MRSA isolates carried PVL genes. Twenty-four (4%) children were BHS carriers. All BHS (n = 8) found in high SES children and six (67%) isolates from slum patients belonged to group A. All group B streptococci were from middle/low SES children, corresponding to five (71%) of the seven BHS isolated in this group. BHS isolates were susceptible to all drugs tested.ConclusionsChildren from different SES communities had distinct bacterial colonization profiles, including MRSA carriage. Public health officials/researchers should consider SES when assessing disease transmission and control measures.

Genotypic and phenotypic β-lactam resistance and presence of PVL gene in Staphylococci from dry bovine udder.

Dairy cows affected with subclinical mastitis can be sources of virulent, antimicrobial-resistant Staphylococci to humans because of the excretion of the bacteria through their milk. This study focussed on the phenotypic and genotypic antibiotic resistance patterns of Staphylococci isolated from dairy cows in early dry period. Among 96 isolates of Gram positive cocci from 157 cows, 76 were identified as Coagulase Negative Staphylococci and the remaining 20 were Staphylococcus aureus. Typical amplicons of coagulase gene were obtained for all 20 samples of S. aureus with three major coagulase types being identified as giving 627 bp (40%), 910 bp (35%) and 710 bp (25%) long PCR products. The groEL gene was amplified in PCR of all 76 isolates of Coagulase Negative Staphylococci, and incubation of PCR products with restriction enzyme PvuII yielded three distinct PCR-RFLP fragment patterns bearing resemblance to S. chromogenes and S. hyicus. Highest sensitivity of Coagulase Negative Staphylococci was noted for Azithromycin (92.5%) and the least to Tetracyclines (76.3%), whereas for S. aureus, it was Cefoperazone (95%) and Azithromycin (72.2%) respectively. Phenotypic resistance to Oxacillin (25 isolates), and Cefoxitin (11 isolates) was detected by dilution method with a commercial strip (Ezy MICTM). Genotypic resistance to β-Lactam antibiotics was found in 65 (34 with mecA gene and 31 with blaZ gene) isolates. Eighteen isolates possessed both the genes, with the PVL gene for virulence being detected in five of them. Nine isolates which had mecA gene were phenotypically susceptible to oxacillin while phenotypic resistance to oxacillin was observed in seven isolates that did not have either mecA or blaZ gene. This is the first report of persistent Staphylococcal infections possessing PVL gene and high level of genotypic resistance to β-Lactam antibiotics in small- holder dairy cattle from India.

Molecular Characterization and Antimicrobial Susceptibility Patterns of Methicillin-Resistant Staphylococcus aureus Isolates in Tabriz, Northwest of Iran

Background: Methicillin resistant Staphylococcus aureus (MRSA) are one of the most common and important pathogens, accounting for diverse nosocomial and community-acquired infections. The serious concern about these bacteria is the development of antibiotic resistance. Objectives: The present study was conducted to investigate the frequency of MRSA strains, their epidemiological and molecular relationships and antibiotic susceptibility patterns of isolated strains from university teaching hospitals of Tabriz, Northwestern Iran (during years 2014 and 2015). Methods: A total of 215 non-repetitive clinical isolates of S. aureus were identified using standard methods. The MRSA isolates were detected by the combination of phenotypic and genotypic methods. The presence of pvl gene and SCCmec types was determined by PCR and multiplex PCR, respectively. The MRSA isolates, in which the presence of mecA gene had been confirmed by PCR, were subjected to Rep-PCR analysis. Resistance to antibacterial agents was determined by disk diffusion, screening agar, and E-test methods. Results: All S. aureus isolates were positive for nuc gene and 87 (40.5%) of them revealed the presence of mecA gene, confirming them as MRSA. All isolates were found to be sensitive to linezolid and vancomycin. However, a reduced sensitivity of 3 MRSA isolates to vancomycin was observed (MIC = 6 μg/mL). SCCmec type III was the most prevalent (79.31%), followed by type IVd (13.80%) and type I (6.90%). The PVL occurrence was detected in 33 (15.35%) S. aureus isolates. The MRSA isolates could be divided to 2 main clusters, indicating the possible clonal relatedness of MRSA isolates. Conclusions: The MRSA isolates with SCCmec type III were the predominant MRSA strains in this area. The majority of MRSA isolates were MDR. Linezolid and vancomycin were found as suitable antibiotics for the treatment of MRSA. The results of typing methods indicated possible clonal relatedness among MRSA isolates. Therefore, routine infection control surveillance is necessary for the prevention of epidemic emergence.

Molecular Detection of Panton - Valentine Leukocidin (PVL) and Methicillin Resistance in Staphylococcus Aureus Pathogen

Background: Methicillin resistance in S. aureus is caused by the acquisition of mecA gene, that encodes an additional s-lactam-resistant penicillin-binding protein, termed PBP2a. PVL toxin is one of many toxins produced by S.aureus. Objectives: to evaluate the efficacy of phenotypic methods and detection of mec A gene with PCR for detection of MRSA and to assess the incidence of PVL gene in MRSA and all S. aureus isolates Methods: 576 patients from Assiut University Hospitals were enrolled in this study were classified into community acquired infection group (CAI) and Hospital acquired infection group (HAI)culture and PCR were done for all samples Results: 92 s.aureus isolates detected MRSA were 62 (67.4%) of all S.aureus infections, They were 30 (65.2 %) of CAI and 32 (69.6 %) of HAI, The prevalence of PVL genes, The prevalence of PVL genes in CAI isolates was 10.9%. None of HAI isolates had PVL gene. Conclusion: The presence of PVL gene cannot be used as a sole marker for CA-MRSA and further studies are required to find a reliable marker or combination of markers to facilitate the recognition of CA- MRSA strains.

Molecular characterization and antimicrobial susceptibility of the CA-MRSA isolated from healthcare workers, Tehran, Iran

Methicillin resistant Staphylococcus aureus (MRSA) has become as a nosocomial pathogen worldwide. Considering the importance of MRSA typing for understanding the evolution and dissemination of these strains, we studied the molecular characteristics of MRSA colonized healthcare workers (HCWs). All MRSA isolated from HCWs, were genotyped using staphylococcal cassette chromosome mec (SCCmec) with multiplex PCR assay, multilocus sequence typing (MLST) and spa typing. Then antibiotic susceptibility pattern and presence of pvl genes were evaluated in MRSA isolates. Cluster analysis by eBURSTv3 showed that MRSA isolates belonged to two major clonal complexes (CC); CC88 (ST88, ST825, ST859) and CC30 (ST39, ST2, ST24) and five singletons. The most prevalent SCCmec type was type IV (70.59%) followed by type V (29.41%). Totally 11 different spa types were discriminated among which type t186 was predominant. All of the MRSA tested (100%) were susceptible to teicoplanin, linezolid and fusidic acid. Totally 52.94% of isolates were positive for pvl genes. The ST88-MRSA-IV accounted for most colonized MRSA isolates. We documented a different molecular epidemiology of MRSA nasal colonization in hospitals under studied, due to the introduction of epidemic clones (ST88, ST39, ST2235, ST80, ST813, ST398, ST825, ST24, ST22, ST859 and ST2).

The Association of Panton-Valentine leukocidin and mecA Genes in Methicillin-Resistant Staphylococcus aureus Isolates From Patients Referred to Educational Hospitals in Ahvaz, Iran.

Background:Staphylococcus aureus, an important human pathogen is one of the main causative agents of nosocomial infection. Virulence genes play a major role in the pathogenicity of this agent and its infections. Methicillin-Resistant Staphylococcus aureus (MRSA) isolates are major challenge among infectious agents that can cause severe infections and mortality. Methicillin-resistant S. aureus produces a unique type of Penicillin Binding Protein 2a (PBP2a) that has low affinity for β-lactam antibiotics. Most of the MRSA bacterial strains can also produce a leukotoxin as Panton-Valentine Leukocidin (PVL) that increases the virulence of MRSA strains and can cause severe necrotic pneumonia. The presence of pvl gene is a genetic marker for the MRSA populations.Objectives:The aim of this study was to explore the association of pvl and mecA genes in clinical isolates of MRSA.Materials and Methods:Fifty MRSA isolates were collected from 200 clinical samples from three different educational hospitals in Ahvaz, Iran, and identified by biochemical tests including catalase, oxidase, tube coagulase, mannitol fermentation, and sensitivity to furazolidone, resistance to bacitracin, PYR test and Voges-Proskauer test. Their resistance to methicillin was evaluated using the disc diffusion method. DNA was extracted by boiling and then the presence of pvl and mecA genes was investigated by the polymerase chain reaction method using specific primers.Results:The results revealed that mecA and pvl genes were positive for 15 (30%) and 3 (6%) of the isolates, respectively. None of mecA positive isolates was positive for pvl gene.Conclusions:It can be concluded from these results that fortunately the prevalence of pvl gene is low in MRSA isolates in this region and there is no association between the presence of pvl and mecA genes in these isolates.

Molecular characterization of methicillin-resistant Staphylococcus aureus: characterization of major clones and emergence of epidemic clones of sequence type (ST) 36 and ST 121 in Tehran, Iran.

Information about the molecular structure of MRSA strains provides significant insights into the epidemiology of this important pathogen. To investigate the molecular characteristics of MRSA isolates, MRSA isolates were subjected to molecular typing by means of spa typing, multilocus sequence typing, Staphylococcal Cassette Chromosome mec (SCCmec) grouping and to phenotypic antimicrobial susceptibility testing by means of disk diffusion assay. Then the presence of pvl genes was evaluated. Cluster analysis by eBURSTv3 showed that MRSA isolates belonged to two major clonal complexes (CC); CC8 (ST239, ST585, ST2732, ST1294) and CC30 (ST30, ST36, ST1163) and four singletons. Subsequent analysis of MRSA isolates revealed that the most prevalent SCCmec type was type III (55.8%) followed by type IV (34.9%) and type II (2.3%). Totally 11 different spa types were discriminated among which types t037 and t030 were predominant. The prevalence of Panton-Valentine leukocidin (PVL)-positive MRSA strains was high (20%), which is a matter of great concern, because the PVL is frequently associated with severe and recurrent SSTIs. ST239-III- t037 represented the most predominant MRSA clone. The presence of sequence type (ST) 36 and ST 121 are being reported for the first time in Iran.

Epidemiology of MRSA in southern Sweden: strong relation to foreign country of origin, health care abroad and foreign travel

All notified MRSA cases in Skåne County have been followed since 2000. We have investigated the MRSA epidemiology over time, method of acquisition, whether some spa types are more prone to spread, and/or cause more infections, and the connection between spa type and country of acquisition/origin. All cases between 2000 and 2010 were included. Infection or colonization and the presence of PVL genes were noted. The spa types of the index cases were correlated with community or healthcare acquisition, proportion of MRSA-positive household contacts, country of origin of families and country of acquisition of MRSA. The number of cases increased from 31 in 2000 to 315 in 2010. Most cases were community-acquired and the median age was 30 years. Thirty-two per cent of the MRSA cases were found because of a clinical infection. Of the household contacts 35 % were MRSA-positive. Only 24 % of the MRSA cases were both of Swedish origin and had contracted MRSA in Sweden. An association between spa type and certain regions of acquisition/origin was noted. Spa types t044, t002 and t008 were the most predominant strains. PVL-positive spa types t008, t019 and t044 caused more skin infections than the other spa types. Our results support screening for MRSA in patients with health care contacts abroad, culturing of patients with skin infections contracted outside Sweden and performing contact tracing among household members. Knowledge of spa type might give guidance in the process of contact tracing. Eradication treatment of MRSA spa types causing more skin infections may be warranted.

A Typical Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Clone Is Widespread in the Community in the Gaza Strip

Epidemiological data on community acquired methicillin-resistant-Staphylococcus aureus (CA-MRSA) carriage and infection in the Middle-East region is scarce with only few reports in the Israeli and Palestinian populations. As part of a Palestinian-Israeli collaborative research, we have conducted a cross-sectional survey of nasal S. aureus carriage in healthy children and their parents throughout the Gaza strip. Isolates were characterized for antibiotic susceptibility, mec gene presence, PFGE, spa type, SCCmec-type, presence of PVL genes and multi-locus-sequence-type (MLST). S. aureus was carried by 28.4% of the 379 screened children-parents pairs. MRSA was detected in 45% of S. aureus isolates, that is, in 12% of the study population. A single ST22-MRSA-IVa, spa t223, PVL-gene negative strain was detected in 64% of MRSA isolates. This strain is typically susceptible to all non-β-lactam antibiotics tested. The only predictor for MRSA carriage in children was having an MRSA carrier-parent (OR = 25.5, P = 0.0004). Carriage of the Gaza strain was not associated with prior hospitalization. The Gaza strain was closely related genetically to a local MSSA spa t223 strain and less so to EMRSA15, one of the pandemic hospital-acquired-MRSA clones, scarcely reported in the community. The rapid spread in the community may be due to population determinants or due to yet unknown advantageous features of this particular strain.

Community-Acquired Staphylococcus aureus Infections in Term and Near-Term Previously Healthy Neonates

Community-acquired, methicillin-resistant Staphylococcus aureus infections are increasing among children. Our goal is to describe the clinical presentation of neonatal community-acquired S aureus disease and provide molecular analyses of the infecting isolates. We retrospectively reviewed the demographics and hospital course of term and near-term previously healthy neonates, < or = 30 days of age, with community-acquired S aureus infections presenting after nursery discharge between August 2001 and March 2005 at Texas Children's Hospital. Prospectively collected isolates were characterized by pulsed-field gel electrophoresis, staphylococcal cassette chromosome mec type, and the presence of PVL genes. Of 89 S aureus infections, 61 were methicillin-resistant S aureus; S aureus infections increased each year. Methicillin-resistant S aureus infections increased from 10 of 20 to 30 of 36 infections from 2002 to 2004. Most subjects, 65 of 89, were male. Symptoms began at 7 to 12 days of age for 26 of 45 male infants with methicillin-resistant S aureus. Most infections, 77 of 89, involved skin and soft tissue; 28 of 61 methicillin-resistant S aureus versus 7 of 28 methicillin-susceptible S aureus infections required drainage. Invasive manifestations included shock, musculoskeletal and urinary tract infection, perinephric abscess, bacteremia, empyema/lung abscess, and a death. Maternal S aureus or skin-infection history occurred with 13 of 61 methicillin-resistant S aureus versus 1 of 28 methicillin-susceptible S aureus infections. The predominant community clone, USA300 (PVL genes +), accounted for 55 of 57 methicillin-resistant S aureus and 3 of 25 methicillin-susceptible S aureus isolates. Community-acquired methicillin-resistant S aureus is a substantial and increasing proportion of S aureus infections in previously healthy neonates. Male infants 7 to 12 days of age are affected most often. Neonatal community-acquired S aureus infection may be associated with concurrent maternal infection. USA300 is the predominant clone among these neonatal isolates in our region.