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1.
Int J Mol Sci ; 23(1)2021 Dec 29.
Article in English | MEDLINE | ID: mdl-35008761

ABSTRACT

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.


Subject(s)
Inflammation/pathology , Leukocidins/metabolism , Mastitis/microbiology , Single-Chain Antibodies/immunology , Staphylococcal Infections/microbiology , Staphylococcus aureus/physiology , Animals , Cattle , Cell Death , Cytokines/metabolism , Female , HEK293 Cells , Humans , Inflammation Mediators/metabolism , Leukocidins/isolation & purification , Mammary Glands, Animal/pathology , Mice , Mice, Inbred BALB C , Neutrophils/metabolism , Peptide Library , Peroxidase/metabolism , Receptors, Chemokine/metabolism , Single-Chain Antibodies/isolation & purification , Staphylococcal Infections/pathology , Staphylococcus aureus/growth & development
2.
J Microbiol Immunol Infect ; 52(2): 242-247, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30181097

ABSTRACT

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.


Subject(s)
Health Personnel , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Nasal Cavity/microbiology , Adult , Bacterial Toxins/isolation & purification , Bacterial Typing Techniques , Carrier State , Congresses as Topic , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Exotoxins/isolation & purification , Female , Genes, Bacterial/genetics , Genotype , Humans , Leukocidins/isolation & purification , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Typing , Multilocus Sequence Typing , Staphylococcal Infections/microbiology , Staphylococcal Infections/transmission , Staphylococcal Protein A/genetics , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification
3.
BMJ Case Rep ; 20182018 Jun 05.
Article in English | MEDLINE | ID: mdl-29871957

ABSTRACT

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.


Subject(s)
Abscess/microbiology , Bacterial Toxins/biosynthesis , Exotoxins/biosynthesis , Leukocidins/biosynthesis , Osteomyelitis/microbiology , Pneumonia, Staphylococcal/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/metabolism , Abdomen/diagnostic imaging , Abscess/complications , Abscess/diagnostic imaging , Abscess/surgery , Adolescent , Anti-Bacterial Agents/therapeutic use , Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Humans , Ilium/diagnostic imaging , Leukocidins/isolation & purification , Lung/diagnostic imaging , Magnetic Resonance Imaging , Male , Methicillin Resistance , Microscopy, Acoustic , Osteomyelitis/complications , Osteomyelitis/diagnosis , Pneumonia, Staphylococcal/complications , Pneumonia, Staphylococcal/diagnosis , Staphylococcal Infections/complications , Staphylococcal Infections/drug therapy , Staphylococcus aureus/drug effects , Staphylococcus aureus/isolation & purification
4.
Am J Med Sci ; 351(3): 279-85, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26992257

ABSTRACT

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.


Subject(s)
Athletes , Nasal Cavity/microbiology , Soccer , Staphylococcus aureus/isolation & purification , Virulence Factors/isolation & purification , Adolescent , Adult , Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Humans , Leukocidins/isolation & purification , Male , Penicillin-Binding Proteins/isolation & purification , Young Adult
5.
Trop Med Int Health ; 21(5): 610-8, 2016 May.
Article in English | MEDLINE | ID: mdl-26970318

ABSTRACT

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.


Subject(s)
Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Penicillin-Binding Proteins/isolation & purification , Staphylococcal Infections/microbiology , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Drug Resistance, Multiple, Bacterial , Exotoxins/genetics , Genetic Carrier Screening/methods , Humans , Indonesia , Leukocidins/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Microbial Sensitivity Tests , Multicenter Studies as Topic , Multilocus Sequence Typing , Multiplex Polymerase Chain Reaction , Penicillin-Binding Proteins/genetics , Spectrum Analysis, Raman , Staphylococcal Infections/genetics , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification , Tertiary Healthcare/statistics & numerical data
6.
Zhonghua Liu Xing Bing Xue Za Zhi ; 37(2): 273-80, 2016 Feb.
Article in Chinese | MEDLINE | ID: mdl-26917530

ABSTRACT

OBJECTIVE: To study the molecular-biologic characteristics and epidemiological status of iatrogenic related Community-acquired methicillin-resistant Staphylococcus (S.) aureus (CA-MRSA) in China through Meta-analysis. METHODS: Data through systematic searching for peer-reviewed articles published before December 3(rd), 2015 from 4 main electronic databases including China National Knowledge Infrastructure (CNKI), Wanfang Data, PubMed and Web of Science Core Collection was collected, for this Meta-analysis. PRISMA guidelines were followed and the proportion of MRSA, CA-MRSA, hospital-acquired MRSA (HA-MRSA) and panton-valentine leucocidin (PVL) gene in certain populations were quantitatively analyzed by Stata 13.0 software. RESULTS: Average proportion of CA-MRSA from S. aureus was 12% (95%CI: 8%-16%). CA-MRSA in MRSA was 18% (95%CI: 12%-24%). 42.1% (95%CI: 20.4%-63.7%) of the CA-MRSA carried a PVL gene, and the number was higher than general MRSA (t=-2.99,P=0.011). CONCLUSION: CA-MRSA was in lower proportion than HA-MRSA, both seen in general MRSA and in S. aureus, but under higher proportion of carrying the PVL gene. Transmission of CA-MRSA could be prevented within the general population through conducting effective surveillances and preventive programs.


Subject(s)
Community-Acquired Infections/microbiology , Cross Infection/microbiology , Iatrogenic Disease/epidemiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/microbiology , Bacterial Toxins/isolation & purification , China/epidemiology , Community-Acquired Infections/epidemiology , Cross Infection/epidemiology , Databases, Factual , Exotoxins/isolation & purification , Humans , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/epidemiology , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification
7.
Pediatr Dev Pathol ; 19(1): 80-5, 2016.
Article in English | MEDLINE | ID: mdl-26367063

ABSTRACT

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.


Subject(s)
Bacterial Toxins/isolation & purification , Coinfection , Enterovirus D, Human/isolation & purification , Enterovirus Infections/virology , Exotoxins/isolation & purification , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Respiratory Tract Infections/microbiology , Respiratory Tract Infections/virology , Staphylococcal Infections/microbiology , Autopsy , Cause of Death , Child , Enterovirus Infections/diagnosis , Enterovirus Infections/therapy , Fatal Outcome , Female , Humans , Predictive Value of Tests , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/therapy , Staphylococcal Infections/diagnosis , Staphylococcal Infections/therapy
8.
PLoS One ; 10(9): e0137531, 2015.
Article in English | MEDLINE | ID: mdl-26348037

ABSTRACT

Few reports from Africa suggest that resistance pattern, virulence factors and genotypes differ between Staphylococcus aureus from nasal carriage and clinical infection. We therefore compared antimicrobial resistance, selected virulence factors and genotypes of S. aureus from nasal carriage and clinical infection in Southwest Nigeria. Non-duplicate S. aureus isolates were obtained from infection (n = 217) and asymptomatic carriers (n = 73) during a cross sectional study in Lagos and Ogun States, Nigeria from 2010-2011. Susceptibility testing was performed using Vitek automated systems. Selected virulence factors were detected by PCR. The population structure was assessed using spa typing. The spa clonal complexes (spa-CC) were deduced using the Based Upon Repeat Pattern algorithm (BURP). Resistance was higher for aminoglycosides in clinical isolates while resistances to quinolones and tetracycline were more prevalent in carrier isolates. The Panton-Valentine leukocidin (PVL) was more frequently detected in isolates from infection compared to carriage (80.2 vs 53.4%; p<0.001, chi2-test). Seven methicillin resistant S. aureus isolates were associated with spa types t002, t008, t064, t194, t8439, t8440 and t8441. The predominant spa types among the methicillin-susceptible S. aureus isolates were t084 (65.5%), t2304 (4.4%) and t8435 (4.1%). spa-CC 084 was predominant among isolates from infection (80.3%, n = 167) and was significantly associated with PVL (OR = 7.1, 95%CI: 3.9-13.2, p<0.001, chi2-test). In conclusion, PVL positive isolates were more frequently detected among isolates from infection compared to carriage and are associated with spa-CC 084.


Subject(s)
Bacterial Toxins/genetics , Exotoxins/genetics , Leukocidins/genetics , Methicillin Resistance/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/genetics , Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Humans , Leukocidins/isolation & purification , Methicillin/therapeutic use , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Microbial Sensitivity Tests , Nigeria/epidemiology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology
9.
Immunol Lett ; 167(1): 47-53, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26141623

ABSTRACT

Vibrio vulnificus hemolysin A (VvhA) is a pore forming toxin and plays an important role in the pathogenesis. The hemolytic and cytotytic property of VvhA toxin is associated with N-terminal leukocidin domain which triggers apoptotic signaling cascade in epithelial cells. The present study was undertaken to assess the protective efficacy of recombinant VvhA leukocidin domain (rL/VvhA) against VvhA toxin challenge using in vitro and in vivo assays. The rL/VvhA protein was found to be non-toxic with no significant hemolytic or cytotoxic effects. Intraperitoneal (I.P.) immunization of BALB/c mice with rL/VvhA protein elicited significantly higher specific serum antibody titer with mixed Th1/Th2 mediated immune responses. HeLa cell monolayer supplemented with anti-rL/VvhA antibodies were effectively protected (viability 86.69%) against lethal 5 LD50 toxin challenge. An effective in vitro proliferation of lymphocyte was observed upon re-stimulation of rL/VvhA primed splenocytes with formalin inactivated VvhA toxin (fVvhA). Co-expression of Th1/Th2 polarized cytokines (IFN-γ, IL-12 and IL-4), were seen in the cell culture supernatant. In contrast to sham immunized mice, rL/VvhA immunized mice demonstrated significant protection (90% survival) against native toxin challenge in vitro and in vivo infection models. These results suggested leukocidin domain of the VvhA toxin as protective immunogen for possible protection against V. vulnificus VvhA.


Subject(s)
Leukocidins/immunology , Protein Interaction Domains and Motifs/immunology , Recombinant Proteins , Toxoids/immunology , Vibrio vulnificus/immunology , Animals , Antibodies, Bacterial/immunology , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Bacterial Proteins/isolation & purification , Cloning, Molecular , Cytokines/metabolism , Female , Gene Expression , Immunization , Leukocidins/genetics , Leukocidins/isolation & purification , Lymphocyte Activation/immunology , Lymphocytes/immunology , Lymphocytes/metabolism , Mice , Mice, Inbred BALB C , Neutralization Tests , Protein Interaction Domains and Motifs/genetics , Spleen/cytology , Spleen/immunology , Spleen/metabolism , Toxoids/genetics , Toxoids/isolation & purification , Vibrio Infections/immunology , Vibrio Infections/mortality , Vibrio Infections/prevention & control , Vibrio vulnificus/genetics
10.
BMC Res Notes ; 7: 862, 2014 Dec 01.
Article in English | MEDLINE | ID: mdl-25435087

ABSTRACT

BACKGROUND: Brain abscess are uncommon childhood infection. Brain abscess caused by Panton-Valentine Leukocidin positive Community acquired Methicillin Resistant Staphylococcal aureus have never been reported in the United Kingdom. CASE PRESENTATION: We report a case of a previously well 11-month old boy of Indian origin who developed a parietal lobe abscess from PVL positive CA-MRSA. CONCLUSION: This case is one of the few described cases of brain abscess caused by PVL CA-MRSA in children. The unusual (insidious) presentation, the absence of a clear staphylococcal focus and the unexpected finding of a CA-MRSA in this patient highlight the challenges of managing such cases in clinical settings and the potential future risk to public health.


Subject(s)
Bacterial Toxins/isolation & purification , Brain Abscess/pathology , Exotoxins/isolation & purification , Leukocidins/isolation & purification , Parietal Lobe/pathology , Staphylococcal Infections/pathology , Anti-Bacterial Agents/therapeutic use , Brain Abscess/drug therapy , Brain Abscess/microbiology , Brain Abscess/surgery , Community-Acquired Infections , Humans , Infant , Male , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/growth & development , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Parietal Lobe/drug effects , Parietal Lobe/microbiology , Parietal Lobe/surgery , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcal Infections/surgery , Treatment Outcome , United Kingdom
11.
J Egypt Public Health Assoc ; 89(3): 114-8, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25534175

ABSTRACT

BACKGROUND AND AIM: Colonization with methicillin-resistant Staphylococcus aureus (MRSA) increases the risk for subsequent infections with an increased mortality and morbidity. Children were suggested to be a major asymptomatic reservoir for community-associated (CA) MRSA with an ability to quickly spread the MRSA within community. Therefore, the availability of epidemiological and antibiotic susceptibility data of CA-MRSA will be useful for the infection control and management policies. This study aimed to assess the nasal carriage, molecular characteristics and antibiotic susceptibility of MRSA in primary school-aged children from Jordan. PATIENTS AND METHODS: A total of 210 nasal swabs were collected from children aged 6-11 years. Isolated MRSA and its SCCmec typing, Spa type and PVL (Panton-Valentine Leukociden) toxin were identified following culture, biochemical and PCR. Antibiogram was determined by the disc diffusion method. RESULTS: The prevalence of CA-MRSA was 7.1%. Allergic rhinitis and recent antibiotic exposure were the only significant risk factors for MRSA nasal carriage among children. Resistance to erythromycin, trimethoprim-sulfamethoxazole and tetracycline was 33.4, 20 and 13.4%, respectively. All isolates were susceptible to the remaining non-ß-lactam antibiotics used in this study, in particular linezolid and mupirocin. All MRSA isolates were SCCmec type IV and PVL toxin negative and the majority were Spa type t223. CONCLUSION AND RECOMMENDATIONS: This is the first study to assess the MRSA prevalence among children aged 6-11 years in Jordan. The prevalence in community children is within the range compared with other studies in other countries. The antibiogram, SCCmec and Spa types of the isolated MRSA are much similar to what was found previously in Jordan. However, all isolates were PVL toxin negative. The study recommends increasing the public awareness of MRSA and the proper antibiotics dispensing. Future studies to follow-up on the changing epidemiology of the CA-MRSA in Jordan are also recommended.


Subject(s)
Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Child , Cross-Sectional Studies , Drug Resistance, Multiple, Bacterial , Exotoxins/isolation & purification , Female , Humans , Jordan/epidemiology , Leukocidins/isolation & purification , Male , Methicillin-Resistant Staphylococcus aureus/drug effects , Nasal Mucosa/microbiology , Penicillin-Binding Proteins/isolation & purification , Polymerase Chain Reaction , Prevalence , Rhinitis, Allergic , Risk Factors , Staphylococcal Infections/drug therapy
12.
Rinsho Byori ; 62(6): 533-7, 2014 Jun.
Article in Japanese | MEDLINE | ID: mdl-25151771

ABSTRACT

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.


Subject(s)
Bacterial Toxins/genetics , Bacterial Toxins/isolation & purification , Exotoxins/genetics , Exotoxins/isolation & purification , Leukocidins/genetics , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections , Bacterial Toxins/biosynthesis , Exotoxins/biosynthesis , Humans , Leukocidins/biosynthesis , Methicillin-Resistant Staphylococcus aureus/genetics , Polymerase Chain Reaction
13.
J Immunol ; 191(12): 6022-9, 2013 Dec 15.
Article in English | MEDLINE | ID: mdl-24190656

ABSTRACT

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.


Subject(s)
Bacterial Proteins/pharmacology , Bacterial Toxins/pharmacology , Leukocidins/pharmacology , Neutrophils/immunology , Staphylococcus aureus/pathogenicity , Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Cell Membrane Permeability/drug effects , Dose-Response Relationship, Immunologic , Electroporation , Exocytosis/drug effects , Extracellular Space , Humans , Leukocidins/isolation & purification , Neutrophils/drug effects , Neutrophils/ultrastructure , Opsonin Proteins/immunology , Peroxidase/metabolism , Respiratory Burst/drug effects , Signal Transduction/drug effects , Staphylococcal Infections/immunology , Staphylococcus aureus/chemistry , Staphylococcus aureus/immunology , Superoxides/metabolism , Virulence
14.
Foodborne Pathog Dis ; 10(8): 705-10, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23692075

ABSTRACT

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.


Subject(s)
Food Contamination/analysis , Meat/microbiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Toxins/genetics , Bacterial Toxins/isolation & purification , Cattle , Chickens , China , Drug Resistance, Multiple, Bacterial , Enterotoxins/genetics , Enterotoxins/isolation & purification , Exotoxins/genetics , Exotoxins/isolation & purification , Food Microbiology , Food Storage , Hong Kong , Leukocidins/genetics , Leukocidins/isolation & purification , Microbial Sensitivity Tests , Swine
15.
New Microbiol ; 36(1): 49-55, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23435815

ABSTRACT

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.


Subject(s)
Bacterial Toxins/genetics , Exotoxins/genetics , Leukocidins/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/genetics , Adolescent , Algeria , Bacterial Toxins/isolation & purification , Bacterial Typing Techniques , Child , Child, Preschool , Community-Acquired Infections/drug therapy , Community-Acquired Infections/microbiology , Cross Infection/drug therapy , Cross Infection/microbiology , Electrophoresis, Gel, Pulsed-Field , Exotoxins/isolation & purification , Humans , Infant , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Microbial Sensitivity Tests , Molecular Epidemiology , Multilocus Sequence Typing , Polymerase Chain Reaction , Staphylococcal Infections/microbiology
16.
Methods Mol Biol ; 943: 105-13, 2013.
Article in English | MEDLINE | ID: mdl-23104284

ABSTRACT

The control and prevention of meticillin-resistant Staphylococcus aureus (MRSA) is a major challenge for healthcare establishments, especially as this pathogen continues to evolve. The emergence and spread of community associated MRSA producing Panton-Valentine leukocidin (PVL) causing severe, sometimes fatal, infections in otherwise healthy people is a significant cause of concern. Patient screening to detect MRSA is now widely used as part of an effective control program to limit the spread of this pathogen. Real-time PCR targeting specific MRSA markers offers a rapid alternative to conventional methods enabling earlier intervention, such as patient isolation and decolonization treatment. Herein we describe a multiplex real-time assay that combines primers and probes to detect MRSA and the genes for PVL to provide a rapid and informative assay.


Subject(s)
Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Leukocidins/isolation & purification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/methods , Bacterial Toxins/genetics , DNA, Bacterial/isolation & purification , Exotoxins/genetics , Humans , Leukocidins/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Nucleic Acids/isolation & purification
17.
Med Princ Pract ; 22(3): 245-9, 2013.
Article in English | MEDLINE | ID: mdl-23147072

ABSTRACT

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.


Subject(s)
Bacterial Toxins/isolation & purification , Exotoxins/isolation & purification , Leukocidins/isolation & purification , Staphylococcus aureus/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Bacteriological Techniques , DNA, Bacterial/genetics , Exotoxins/genetics , Female , Humans , Kuwait/epidemiology , Leukocidins/genetics , Male , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Penicillin-Binding Proteins , Respiratory Tract Infections/microbiology , Staphylococcal Skin Infections/microbiology , Staphylococcus aureus/enzymology , Staphylococcus aureus/genetics
18.
Foodborne Pathog Dis ; 9(10): 962-5, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22953738

ABSTRACT

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.


Subject(s)
Staphylococcal Infections/epidemiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/pathogenicity , Swine Diseases/epidemiology , Swine/microbiology , Adult , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/isolation & purification , Bacterial Toxins/isolation & purification , Bacterial Toxins/metabolism , Cloning, Molecular , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Exotoxins/isolation & purification , Exotoxins/metabolism , Female , Gene Expression Profiling , Genotype , Humans , Leukocidins/isolation & purification , Leukocidins/metabolism , Male , Methicillin/pharmacology , Methicillin Resistance , Middle Aged , Multilocus Sequence Typing/veterinary , Penicillin-Binding Proteins , Senegal/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification , Swine Diseases/microbiology
19.
Am J Primatol ; 74(12): 1071-5, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22907634

ABSTRACT

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.


Subject(s)
Animals, Zoo/microbiology , Drug Resistance, Bacterial , Pan troglodytes/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/isolation & purification , Animals , Bacterial Toxins/isolation & purification , Cross-Sectional Studies , Endangered Species , Exotoxins/isolation & purification , Genotype , Humans/microbiology , Leukocidins/isolation & purification , Staphylococcal Infections/transmission , Staphylococcus aureus/chemistry , Staphylococcus aureus/physiology
20.
J Ocul Pharmacol Ther ; 28(1): 12-6, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22014156

ABSTRACT

PURPOSE: Staphylococcus aureus (SA) endophthalmitis is generally a postsurgical infection with an undefined source of entry. Hospital-acquired (HA) SA infections are associated with multi-antibiotic resistance and absence of the Panton-Valentine Leukocidin (PVL) toxin. Community-acquired (CA) SA infections are not associated with multi-antibiotic resistance and possess the PVL toxin. We hypothesize that CA infection is more common than HA for SA endophthalmitis. METHODS: Twenty de-identified SA isolates, collected from the vitreous and/or aqueous of clinical endophthalmitis, were tested for the presence of PVL toxin and antibiotic susceptibility. PVL testing was performed using a kit to detect the Staphylococcal toxin by reversed passive latex agglutination (PVL-RPLA "Seiken," Denka Seiken Co., LTD). SA isolates were tested for antibiotic susceptibility using disk diffusion at the time of isolation. Multi-antibiotic resistance was defined as resistance to at least 3 classes of antibiotics. RESULTS: Of the 20 isolates, 15 were multi-antibiotic resistant and PVL-negative consistent with HA, and 1 was not multi-antibiotic resistant and PVL-positive, consistent with CA. Two isolates tested positive for PVL with one demonstrating both methicillin and fluoroquinolone (FQ) resistance. Of the 18 PVL-negative SA isolates, 15 (83%) were multi-antibiotic resistant (12 methicillin-resistant SA, 14 FQ resistant). CONCLUSIONS: Our results reject the hypothesis that SA isolated from endophthalmitis is consistent with CA sources due to the lack of the PVL toxin and multiple resistant patterns of the SA. PVL does not appear to be a key virulence factor for the development of SA endophthalmitis.


Subject(s)
Cross Infection/microbiology , Endophthalmitis/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Toxins/isolation & purification , Community-Acquired Infections/drug therapy , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Cross Infection/drug therapy , Cross Infection/epidemiology , Drug Resistance, Multiple, Bacterial , Endophthalmitis/drug therapy , Endophthalmitis/epidemiology , Exotoxins/isolation & purification , Humans , Leukocidins/isolation & purification , Microbial Sensitivity Tests , Staphylococcal Infections/drug therapy , Staphylococcal Infections/epidemiology , Staphylococcus aureus/drug effects
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