Ability of biofilm production and molecular analysis of spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus (original) (raw)
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FEMS Immunology & Medical Microbiology, 2007
Staphylococcus aureus is one of the most important etiological agents responsible for healthcare-associated infections and is capable of producing many virulence factors including biofilm. The aim of the present study was to analyze the correlation between the presence of the icaD and icaA genes and the ability to produce biofilm in vitro in 302 methicillin-resistant (MRSA) and 268 methicillinsensitive S. aureus (MSSA) strains isolated in the Provincial Hospital in Gdansk. Presence of the icaD and icaA genes was detected by PCR and the ability to produce biofilm in vitro was measured both spectrophotometrically and via Congo Red Agar plate culture methods. We found that 91% of MRSA strains harbored the icaD gene. Moreover, all icaD-negative strains were icaA-positive. Of MRSA and MSSA strains, 47% and 69%, respectively, produced biofilm in vitro. The level of consistency between the two applied phenotypic methods was 96%. Additionally, we found that strains with the same biofilm status may be present in asymptomatic carriers and cause infections.
BMC Infectious Diseases, 2018
Background: Methicillin resistant Staphylococcus aureus (MRSA) is recognized worldwide as a leading cause of hospital and community infections. Biofilm formation by MRSA is an extremely important virulence factor to be understood. Our aim was to establish phenotypic and genotypic characterization of virulence factors among 43 MRSA clinical isolates in a Tunisian hospital. Methods: We investigated enzymatic profiles, biofilm production and prevalences of genes encoding intracellular adhesion molecules (icaA and icaD), Microbial Surface Components Recognizing Adhesive Matrix Molecules genes (fnbA, fnbB and cna) and exoenzymes genes (geh, sspA and sspB). Results: Our findings revealed that caseinase, gelatinase, lipase and lecithinase activities were detected in 100%, 100%, 76.6% and 93.3% of cases respectively. This study showed that 23 strains (76.7%) were slime producers on Congo red medium. Furthermore, 46.5% and 53.5% of isolates were respectively highly and moderately biofilm-forming on polystyrene. Significant association was found between both biofilm tests. PCR detection showed that 74.4%, 18.6%, 69.8%, 65.1% and 74.4% of isolates harbored fnbA, fnbB, icaA, icaD and cna genes respectively. In addition, 34.9%, 18.6% and 30.2% of MRSA strains were found positive for sspA, sspB and geh genes respectively. Further, statistical data showed that the presence of the fnbA and fnbB genes was significantly associated with a high biofilm production on polystyrene. However, no statistical association was observed for the icaA, icaD and cna genes. Conclusions: This study indicates that the detection of fnbA and fnbB contributing to the first step of biofilm formation has been predictable of high biofilm production. As studied factors contribute to MRSA virulence, this research could be of value in orienting towards the development of new preventive and therapeutic measures.
Folia Medica Indonesiana, 2018
Methicillin-resistant Staphylococcus aureus (MRSA) is a global health concern. One of the factors causing hospital infection is related to the ability of MRSA bacteria to form biofilms. Polysaccharide intercellular adhesin (PIA), encoded by ica gene, have an important role in S. aureus intracellular accumulation and aggregation. The aims of this study was to analyze the relationship between icaA, icaD genes and biofilm production in MRSA carrier and clinical isolate in Dr. Soetomo Hospital Surabaya. This study was an observational study using cross sectional approach. The sample was 47 MRSA isolates is as follow 28 isolates from carrier and 19 were clinical isolates. All of MRSA isolates carried mecA gene. PCR was performed to detect icaA and icaD genes. Biofilm formation was detected using microtiter plate assay (MTP). icaA gene was detected in all isolates whereas icaD gene in 96,4% carrier isolates and all (100%) of clinical isolates. Positive MTP results showed in all (100%) of carrier isolates and 57,9% of clinical isolates. Statistic result was significantly different in biofilm formation between carrier and clinical MRSA isolates. The proportion of positive biofilm formation in isolate with positive icaA/D genes was 82.6%. There was not any association between icaA and icaD gene with biofilm production.
Biofilm Formation and Methicillin Resistance of Staphylococcus aureus Isolated from Clinical Samples
The International Arabic Journal of Antimicrobial Agents, 2020
Background: Staphylococcus aureus including methicillin resistant S. aureus (MRSA) is one of the most effective biofilm-forming organisms, biofilm contribute in protecting the microorganism from host defenses and prevent the effective penetration of antimicrobial agents. Biofilm formation is considered as an important contributing factor for the initiation and establishment of chronic infection by S. aureus and known as a major obstacle in the treatment of S. aureus infections is their ability to develop resistance to antimicrobials. Aims : To screen clinical Staphylococcus aureus including MRSA isolates for their biofilm forming abilities and their association with antimicrobial resistance. Methods: A total of 196 clinical isolates of S. aureus were obtained from different sample sources using standard microbiological techniques from three major hospitals in Gaza strip. Biofilm formation of these isolates was determined by tissue culture plate (TCP) method and tube adherence method...
Introduction: Biofilm producing Staphylococcus aureus is known as one of the major causative agents of infections, failure of implanted devices and persistent infection among hospitalized patients. The aim of the present study was to determine the frequency of biofilm producing S. aureus isolates amongst the clinical specimens. Methods: This cross-sectional study was conducted during 2012 to 2013 in two teaching hospitals in Shiraz, southwest of Iran. Totally, 345 S. aureus isolates from various clinical specimens were included. Biofilm producing isolates were phenotypically detected using Congo Red Agar (CRA) and genotypically by PCR assay for the icaA and icaD genes. Results: Of the 345 S. aureus isolates, 42.3% were methicillin-resistant S. aureus (MRSA) and subsequently 57.7% were methicillin susceptible isolates. The results of CRA plates showed that 77 (52.7%) and 74 (37.2%) of MRSA and MSSA were biofilm producing isolates. The frequency of icaA/D genes among MRSA and MSSA iso...
2014
Objective: The aims of present study were to determine the antimicrobial susceptibility, virulence factors and biofilm formation among MRSA hospital isolates of Staphylococcus aureus. Methods: Thirty non-repetitive strains of S. aureus isolated from three hospitals in Kerman, Iran. Antimicrobial susceptibility was determined by disk diffusion breakpoints method according to CLSI guideline. The minimum inhibitory concentration (MIC) of vancomycin and methicillin were measured by the broth microdilution and E-test procedures. Virulence factors (protease, DNase, lecithinase, capsule and hemolysis) associated with the above isolates was studied. Biofilm was quantified by microtiter technique. Results: In total, 14 (46.7%) S. aureus were isolated from lower respiratory tract, six (20.0%) from urinary tract and remaining 10 (33.3%) were recovered from wounds, blood and orthopedic patients. All of the isolates were susceptible to tigecycline, eight (26.7%) were found to be resistant to methicillin (MRSA) and 4 (13.3%) showed reduced susceptibility to vancomycin. No any vancomycin resistant isolate was detected (p≤0.05). MIC results showed that four of the isolates (13.3%) exhibited MIC 4 μg/mL to vancomycin while, five (16.6%) demonstrated MIC 32 μg/mL to methicillin. The isolates were also resistant to amoxicillin/clavulanic acid, tetracycline and tobramycin. It was found that, six (75 %) of MRSA strains produced lecithinase, seven (96.7%) demonstrated protease and DNase activities as compared to MSSA isolates. Biofilm analysis revealed that twenty (66.7%) isolates formed strong, seven (23.3%) formed moderate and three (10.0%) had weak biofilm.
Diseases
The increasing incidence of methicillin-resistant and biofilm-forming S. aureus isolates in hospital settings is a gruesome concern today. The main objectives of this study were to determine the burden of S. aureus in clinical samples, assess their antibiotic susceptibility pattern and detect biofilm formation and mecA gene in them. A total of 1968 different clinical specimens were processed to isolate S. aureus following standard microbiological procedures. Antibiotic susceptibility test of the isolates was performed by Kirby–Bauer disc-diffusion method following CLSI guidelines. Biofilm was detected through tissue culture plate method. Methicillin-resistant S. aureus (MRSA) isolates were screened using cefoxitin (30 µg) discs and mecA gene was amplified by conventional polymerase chain reaction (PCR). Of 177 bacterial growth, the prevalence of S. aureus was 15.3% (n = 27). MRSA were 55.6% (15/27) and 44% (12/27) exhibited multidrug resistance (MDR). There was no significant associ...
D. A. Oche, 2020
Biofilm formation and resistance to methicillin are among the factors that makes Staphylococcus aureus a very important human pathogen in both health-care and community settings. This study investigated methicillin-resistance among biofilm-producing S. aureus isolated from 49 orthopaedic in-patients within a 3 months period. Wound swabs, nasal swabs, bed swabs and urine samples were collected from each patient. The samples were cultured and screened for presence of S. aureus while the micro-titre plate method was used to detect biofilm producing isolates. PCR technique was finally used to detect the presence of mecA gene in methicilin resistant S. aureus (MRSA) isolates. Findings reveal 14.8% of bacterial isolates were Staphylococcus aureus of which 96.4% were biofilm-producers. However, strong biofilm producers constitute 11.1%. The mecA gene was detected in 15.8% of the MRSA isolates. Therefore, MRSA among biofilm-producing S. aureus is a potential threat primarily to the community of National Orthopaedic Hospital Dala and a major public health challenge.
BioMed research international, 2013
Periodic monitoring of Staphylococcus aureus characteristics in a locality is imperative as their drug-resistant variants cause treatment problem. In this study, antibiograms, prevalence of toxin genes (sea-see, seg-ser, seu, tsst-1, eta, etb, and etd), PFGE types, accessory gene regulator (agr) groups, and ability to form biofilm of 92 S. aureus Thailand clinical isolates were investigated. They were classified into 10 drug groups: groups 1-7 (56 isolates) were methicillin resistant (MRSA) and 8-10 (36 isolates) were methicillin sensitive (MSSA). One isolate did not have any toxin gene, 4 isolates carried one toxin gene (seq), and 87 isolates had two or more toxin genes. No isolate had see, etb, or tsst-1; six isolates had eta or etd. Combined seg-sei-sem-sen-seo of the highly prevalent egc locus was 26.1%. The seb, sec, sel, seu, and eta associated significantly with MSSA; sek was more in MRSA. The sek-seq association was 52.17% while combined sed-sej was not found. Twenty-three P...
PLOS ONE, 2024
Biofilm development significantly enhances the virulence of methicillin-resistant Staphylococcus aureus (MRSA), leading to severe infections and decreased susceptibility to antibiotics, especially in strains associated with hospital environments. This study examined the occurrence of MRSA, their ability to form biofilms, agr typing, and the antibiotic resistance profiles of biofilm-forming MRSA strains isolated from environmental surfaces at Mymensingh Medical College Hospital (MMCH). From 120 swab samples, 86 (71.67%) tested positive for S. aureus. MRSA was identified in 86 isolates using the disk diffusion technique, and by polymerase chain reaction (PCR), 56 (65.1%) isolates were confirmed to carry the mecA gene. The Crystal Violet Microtiter Plate (CVMP) test revealed that 80.35% (45 isolates) were biofilm-forming and 19.6% (11 isolates) were non-biofilm-forming. Out of 45 biofilm producer isolates 37.5% and 42.9% isolates exhibited strong and intermediate biofilm-forming characteristics, respectively. Molecular analysis revealed that 17.78% of MRSA isolates carried at least one gene related to biofilm formation, specifically icaA, icaB, and icaD genes were discovered in 13.33%, 8.89%, 6.67% of the MRSA isolates, respectively. In agr typing, the most prevalent group was agr I (71.11%), followed by group III (17.78%) and group II (11.11%). Group IV was not detected. The distribution of agr gene groups showed a significant difference among biofilm-forming isolates (p < 0.05). In agr group I, 18.75% of isolates carried the icaA gene, 12.5% carried the icaB gene, and 9.37% carried the icaD gene. Biofilm-forming genes were not detected in any of the isolates from agr groups II or III. There are no statistically significant differences between agr groups and the presence of these genes (p > 0.05). Antibiotic resistance varied significantly among agr groups, with agr group I displaying the highest resistance, agr group II, and agr group III exhibiting the least resistance (p < 0.05). Seventy-three (73.3%) of the isolates were multi-drug resistant, with agr group I displaying nineteen MDR patterns. The occurrence of MRSA in hospital environments and their capacity to form biofilm raises concerns for public health. These findings support the importance of further research focused on agr quorum sensing systems as a basis for developing novel antibacterial agents.