scholarly journals Comparison of mecA gene-based PCR with CLSI cefoxitin and oxacillin disc diffusion methods for detecting methicillin resistance in Staphylococcus aureus clinical isolates

2013 ◽  
Vol 7 (21) ◽  
pp. 2438-2441 ◽  
Author(s):  
Rostami Soodabeh ◽  
Moosavian Mojtaba ◽  
Shoja Saeed ◽  
Torabipour Maryam ◽  
Farshadzadeh Zahra
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Clinical Isolates ◽  
Disc Diffusion

scholarly journals Comparison of Cefoxitin and Oxacillin disc diffusion test for the detection of mecA mediated methicillin resistance in Staphylococcus aureus

2013 ◽  
Vol 7 (1) ◽  
pp. 07-10 ◽  
Author(s):  
Durdana Chowdhury ◽  
Sanya Tahmina Jhora ◽  
Shika Paul ◽  
Tarek Mahbub Khan ◽  
Mili Rani Saha ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Clinical Samples ◽  
Disc Diffusion ◽  
Standard Laboratory ◽  
Diffusion Test ◽  
Laboratory Procedure ◽  
Gold Standard Method

The study was designed to evaluate the efficacy of cefoxitin disc diffusion test to detect methicillin resistance in Staphylococcus aureus and compare it with oxacillin disc diffusion test and detection of mecA gene by PCR.  A total 116 S. aureus were isolated from clinical samples, collected from SSMC&MH, BIRDEM and NMC hospital, and was isolated by culture and identified by standard laboratory procedure. Antibiotic susceptibility testing was performed by oxacillin (1µg) and cefoxitin (30 µg) discs. PCR for amplification of mecA gene was performed as a gold standard method. Out of 116 isolates, 28 were PCR positive, 33 and 31 were oxacillin and cefoxitin resistant respectively. The sensitivity and specificity for the detection of MRSA was 100% and 94.31%inoxacillin disc diffusion test, and 96.42% and 95.45%in cefoxitin disc diffusion test respectively. Specificity is higher (95.45%) in cefoxitin disc diffusion test than oxacillin disc diffusion test in the detection of MRSA. Use of disc diffusion tests for both oxacillin and cefoxitin can help in more accurate prediction of methicillin resistance than single test, especially in centers which are not equipped to carry out more sophisticated tests for the detection of MRSA.DOI: http://dx.doi.org/10.3329/bjmm.v7i1.19314 Bangladesh J Med Microbiol 2013; 07(01): 7-10

scholarly journals Isolation and Host Range of Lytic Staphylophages on Clinical Isolates of Methicillin-Resistant Staphylococcus aureus

2020 ◽  
Vol 11 (3) ◽  
Author(s):  
Mahtab Sadat Madani Boroujeni ◽  
Mohammadreza Mahzounieh ◽  
Azizollah Ebrahimi Kahrizsangi ◽  
Soudabeh Rostami ◽  
Azam Mokhtari ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bacterial Infections ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Plaque Assay ◽  
Clinical Isolates ◽  
Teaching Hospitals ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Methicillin Resistant

Background: Staphylococcus aureus (S. aureus) is a major cause of nosocomial infections in humans and animals. Because of the widespread resistance to antibiotics, microbiologists are trying to find other therapeutic interventions such as phage therapy for bacterial infections. Objectives: The present study aimed to isolate staphylophages with lytic effects on methicillin-resistant S. aureus (MRSA) clinical isolates as a potential alternative agent to antibiotic therapy. Methods: This experimental, descriptive study is performed in the Microbiology Laboratory of Shahrekord University (Iran) from September 2018 to March 2019. Two cocktails of staphylophages were isolated from Isfahan (Iran) urban sewage samples. The double-layer agar method was used to detect lytic phages. Morphology characteristic by transmission electron microscopy (TEM) images was used to identify staphylophages. One hundred and thirty three S. aureus were isolated from clinical samples of two teaching hospitals in Isfahan and Shiraz, Iran. Methicillin resistance and the presence of the mecA gene were determined by the disk diffusion method and polymerase chain reaction (PCR) assay, respectively. The phage susceptibility of mecA positive isolates was determined by plaque assay. Results: Two staphylophage cocktails were prepared, which had lytic effects on forty-four MRSA isolates. Cocktails 1 and 2 lysed 19 (14.2%) and 25 (18.7%) isolates, respectively. Of 133 S. aureus isolates, 88.7% carried the mecA gene. Conclusions: Different bacteriophages in two phage cocktails had relatively good lytic effects on S. aureus clinical isolates. Therefore, phage cocktails may be an appropriate alternative to antibiotics against S. aureus.

scholarly journals Resistance pattern of methicillin resistant Staphylococcus aureus among nasal isolates of HIV infected patients in a tertiary care hospital

2019 ◽  
Vol 7 (2) ◽  
pp. 428
Author(s):  
Manish Kumar Diwakar ◽  
Ankur Goyal ◽  
Sapna Goyal
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Tertiary Care ◽  
Meca Gene ◽  
Nasal Carriage ◽  
Control Group ◽  
Care Hospital ◽  
Disc Diffusion ◽  
Increased Risk ◽  
Nasal Swab Sample

Background: Patients infected with HIV have an increased risk of nasal Staphylococcus aureus carriage as well as consecutive staphylococcal infections and is a major reservoir for MRSA which is potential risk factors for community acquired MRSA. Knowing the Nasal carriage status of Staphylococcus aureus and their Antibiogram will be beneficial for effective management of these patients.Methods: Nasal swab sample were collected from all the participants and processed for culture and identification of Staphylococcus aureus and their antimicrobial sensitivity. All the Staphylococcus aureus isolates were tested for Methicillin resistance by Oxacillin screen agar test, cefoxitin disc diffusion test and further confirmed by mecA gene PCR.Results: In this study out of 220 HIV seropositive patients, 43.64% isolates were confirmed to be S. aureus, 18.75% MRSA and 81.25% were MSSA. Cefoxitin disc diffusion showed 100% specificity (95% CI; 97.05%-100.00%), 100% sensitivity (95% CI; 83.89-100.00%) and 100% accuracy (95% CI; 97.47% to 100.00%) while comparing with gold standard mecA gene PCR. Among the nasal carriers; males (60%) ware dominant on females (40%). 31-50 years age group was strongly associated with MRSA nasal carriage. None of the isolates were resistant against lenozolid, teicoplanin and vancomycin while ampicillin (75%), ciprofloxacin (62.5%), clindamycin (59.38%) and cotrimoxazole (53.13%) showed increased resistance against S. aureus nasal carriage.Conclusions: Resistance among HIV positive persons for all antibiotics showed statistically significant while compared to control group. Cefoxitin disc diffusion can be used as surrogate agent for mecA gene detection.

Assessment of the Multifactorial Effect on Antimicrobial Sensitivity in Positive Staphylococcus Aureus Clinical Isolates from Assir Region, Saudi Arabia

2012 ◽  
Vol 13 (2) ◽  
pp. 152-159 ◽  
Author(s):  
Nazar M Abdalla ◽  
Waleed O Haimour ◽  
Amani A Osman ◽  
Hassan Abdul Aziz
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Culture Media ◽  
Meca Gene ◽  
Laboratory Data ◽  
Age Groups ◽  
Clinical Isolates ◽  
Diabetic Patients ◽  
Staph Aureus ◽  
Antimicrobial Sensitivity

General objectives: This study aimed at assessment of factors affecting antimicrobial sensitivity in Staphylococcus aureus clinical isolates from Assir region, Saudi Arabia. Materials and Methods: In this study, eighty one patients presented with Staph. aureus infections either nosocomial or community acquired infections were involved by collecting nasal swabs from them at Aseer Central Hospital General Lab. These patients were from all age groups and from males and females during the period of Jan 2011- Jun 2011. These samples were undergone variable laboratory procedures mainly; bactech, culture media, antibiotics sensitivity test using diffusion disc test (MIC) and molecular (PCR) for detection of mec A gene. Clinical and laboratory data were recorded in special formats and analyzed by statistical computer program (SPSS). Results: Showed that; Descriptive and analytical statistical analysis were performed and final results were plotted in tables. In Staph aureus MecA gene positive cases (50) showed: Oxacillin/ Mithicillin, Ciprofloxacin and Fusidin resistant in diabetic patients were 13, 26.0%, 9, 18% and 7, 14% respectively and in non diabetic patients were 37, 74.0%, 22, 44% and 20, 40% respectively. While no sensitivity in diabetic and non diabetic patients using Oxacillin/ Mithicillin. In Staph aureus MecA gene negative cases (31) showed: Oxacillin/ Mithicillin, sensitivity in diabetic patients (5, 16.1%) and in non diabetic were (26, 83.9%). While no resistant in diabetic and non diabetic patients. In Ciprofloxacin and Fusidin resistant in diabetic patients were 1, 3.2% and 1, 3.2% respectively and in non diabetic patients were 12, 38.7% and 7, 22.6%respectively. Erythromycin in Staph aureus ( MecA gene) positive cases (50) showed: resistant in age (0-15) years were (5, 10%), (16-50) years were (16, 32%) and ( ›50 years) were (12, 24%). Erythromycin in Staph aureus (MecA gene) negative cases (31) showed: resistant in age (0-15) years were (6, 19.3%), (16-50) years were (5, 16.1%) and ( ›50 years) were (3, 9.7%). Conclusion: Drugs resistance is a major progressive multifactorial problem facing the treatment of Staph aureus infections. DOI: http://dx.doi.org/10.3329/jom.v13i2.12750 J Medicine 2012; 13 : 152-159

scholarly journals Isolation of Biofilm Producing Methicillin-Resistant Staphylococcus aureus from Hospitalized Orthopaedic Patients in Kano State, Nigeria

2021 ◽  
Vol 28 (1) ◽  
pp. 66-74
Author(s):  
D.A. Oche ◽  
U. Abdulrahim ◽  
A.S. Oheagbulem ◽  
B.O. Olayinka
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Community Settings ◽  
Potential Threat ◽  
Orthopaedic Patients ◽  
Nasal Swabs ◽  
Public Health Challenge

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. Keywords: Biofilm, Methicillin-resistance Staphylococcus aureus (MRSA), mecA gene, Orthopaedic patient

scholarly journals Susceptibility of Clinical Isolates of Staphylococcus aureus to Ceftaroline

2021 ◽  
Author(s):  
Harsha Sreedharan ◽  
KB Asha Pai
Keyword(s):  
Staphylococcus Aureus ◽  
Tertiary Care Hospital ◽  
Methicillin Resistance ◽  
Descriptive Analysis ◽  
Tertiary Care ◽  
The United States ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Care Hospital

Introduction: Methicillin-Resistant Staphylococcus aureus(MRSA) infection is a major global healthcare problem, the prevalence of which varies from 25-50% in India. It is known to cause Skin and Soft tissue Infections (SSI), endovascular infections, endocarditis, pneumonia, septic arthritis, osteomyelitis, and sepsis. Vancomycin is the drug of choice for treating severe MRSA infections. Ceftaroline, a fifth-generation cephalosporin has been approved by the United States Food and Drug Administration (US FDA) for treating acute bacterial SSI caused by susceptible micro-organisms including MRSA, Community acquired respiratory tract infection, MRSA bacteremia and endocarditis. Aim: To assess the susceptibility of clinical isolates of S. aureusto ceftaroline, in a Tertiary Care Hospital. Materials and Methods: This prospective study was conducted in the Department of Microbiology of a Tertiary Care Hospital over a period of two months from June 2019 to July 2019. S.aureus isolates from various clinical samples were screened for methicillin resistance by disc diffusion method using cefoxitin disc and ceftaroline susceptibility of these isolates was assessed by E-strip method. The isolates were classified as ceftaroline susceptible, Susceptibility Dose Dependent (SDD) and ceftaroline resistant respectively as per CLSI guidelines. A descriptive analysis of the data was done and the results were presented as frequencies and percentages. Results: All the S.aureus isolates were found to be susceptible to ceftaroline. Methicillin Sensitive Staphylococcus aureus(MSSA) isolates had lower Minimum Inhibitory Concentration (MIC) when compared to MRSA. The highest MIC among MRSA was 0.5 μg/mL. Conclusion: Ceftaroline can be considered as an effective alternative for treatment of infections caused by MRSA.

scholarly journals Detection of mecA gene by latex agglutination and its molecular analysis by PCR in methicillin resistant staphylococcus aureus.

2020 ◽  
Vol 27 (07) ◽  
pp. 1363-1370
Author(s):  
Aneela Khawaja ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Saeed Anwar ◽  
Faiqa Arshad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Medical Institute ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Agglutination Method ◽  
Resistant Strains

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

scholarly journals Prevalence of Methicillin Resistant and Virulence Determinants in Clinical Isolates of Staphylococcus aureus

2018 ◽  
Vol 10 (1) ◽  
pp. 108-115
Author(s):  
Manjunath Chavadi ◽  
Rahul Narasanna ◽  
Ashajyothi Chavan ◽  
Ajay Kumar Oli ◽  
Chandrakanth Kelmani. R
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Morphology ◽  
Methicillin Resistance ◽  
Protein A ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Virulence Determinants ◽  
Methicillin Resistant ◽  
Alternative Medicines ◽  
Mrsa Strains

Introduction:Methicillin-resistantStaphylococcus aureus(MRSA) is the major threat that is a result of the uncontrolled use of antibiotics causing a huge loss in health, so understanding their prevalence is necessary as a public health measure.Objective:The aim of this study was to determine the prevalence of methicillin-resistant MRSA and virulence determinant among associatedS. aureusfrom the clinical samples obtained from various hospital and health care centers of the Gulbarga region in India.Materials and Methods:All the collected samples were subjected for the screening ofS. aureusand were further characterized by conventional and molecular methods including their antibiotic profiling. Further, the response of methicillin antibiotic on cell morphology was studied using scanning electron microscopy.Results:A total 126S. aureuswas isolated from the clinical samples which showed, 100% resistant to penicillin, 55.5% to oxacillin, 75.3% to ampicillin, 70.6% to streptomycin, 66.6% to gentamicin, 8.7% to vancomycin and 6.3% to teicoplanin. The selected MRSA strains were found to possessmecA(gene coding for penicillin-binding protein 2A) andfemA(factor essential for methicillin resistance)genetic determinants in their genome with virulence determinants such as Coagulase (coa) and the X region of the protein A (spa)gene. Further, the methicillin response in resistantS. aureusshowed to be enlarged and malformed on cell morphology.Conclusion:The molecular typing of clinical isolates ofS. aureusin this study was highly virulent and also resistant to methicillin; this will assist health professionals to control, exploration of alternative medicines and new approaches to combat Staphylococcal infections more efficiently by using targeted therapy.

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