scholarly journals MecA gene and methicillin-resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia

2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Sancaka Chasyer Ramandinianto ◽  
Aswin Rafif Khairullah ◽  
Mustofa Helmi Effendi
Keyword(s):  
Staphylococcus Aureus ◽  
Pathogenic Bacteria ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Dairy Farms ◽  
Detection Methods ◽  
Disc Diffusion ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Oxacillin Resistance

Abstract. Ramandinianto SC, Khairullah AR, Effendi MH. 2020. MecA gene and methicillin-resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia. Biodiversitas 21: 3562-3568. Milk Borne Disease (MBD) can be caused by a variety of pathogenic bacteria, one of which is Staphylococcus aureus which has a large impact on aspects of public health. The therapy used to treat staphylococcal infection is Oxacillin preparations that can inhibit bacterial wall synthesis, but the adaptation of the mecA gene to staphylococcal cassette chromosome mec (SCCmec) causes the emergence of strains of methicillin-resistant S. aureus (MRSA). The purpose of this study was to detect the level of MRSA strain contamination in dairy cows in East Java by comparing the mecA gene, Oxacillin, and Cefoxitin Disc Diffusion Methods and Oxacillin Resistance Screen Agar (ORSA) detection methods. A total of 150 cow's milk samples were taken at 3 village dairy farms in East Java, samples were added to the enrichment media Buffer Pepton Water (BPW) and then isolates were planted and purified using Mannitol Salt Agar (MSA). The detection of MRSA was carried out by the Kirby Bauer disc diffusion preparation Cefoxitin 30 μg and Oxacillin 30 μg then confirmed by ORSA and the presence of mecA gene by the polymerase chain reaction (PCR) method. The results showed that from a total of 92 S. aureus isolates using Oxacillin disc test, 24 resistant isolates were obtained, using Cefoxitin disc test, 17 isolates were obtained, and using the ORSA test 18 MRSA isolates were obtained. MRSA isolates tested by PCR obtained evidence of 2 isolates of mecA gene. It can be concluded that the Oxacillin disc test was the highest sensitivity for detecting MRSA strain isolate, however, mecA gene was the golden standard to detect MRSA on the dairy farms.

scholarly journals Dressed chicken as potential vehicle for spread of methicillin-resistant Staphylococcus aureus in Sokoto, Nigeria

2020 ◽  
Vol 6 (10) ◽  
pp. FSO619
Author(s):  
Ibrahim A Musawa ◽  
Yusuf Yakubu ◽  
Bashiru Garba ◽  
Fatimah M Ballah ◽  
Hassan Abdurrahman Jibril ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Resistance Screening ◽  
Methicillin Resistant ◽  
Oxacillin Resistance ◽  
Chicken Carcass ◽  
Biochemical Analyses

Aim: To evaluate the role of dressed chicken in the spread of methicillin-resistant Staphylococcus aureus (MRSA) in Sokoto, Nigeria. Materials and methods: 190 chicken carcass rinsates were subjected to culture and biochemical analyses to isolate and identify MRSA. PCR was used to amplify mecA gene that is responsible for methicillin resistance. Results & conclusion: Culture and molecular analysis showed 19.5% (37/190) of the rinse had MRSA on oxacillin-resistance screening agar base (ORSAB) with 7.9% (15/190) possessing the mecA gene. Significant association (p = 0.044) exist between local-chicken and presence of MRSA, being twice more likely to have MRSA compared to exotic-chickens (odds ratio [OR] = 2.132). Results indicate possible role of dressed-chicken in the spread of MRSA. Authorities should regulate the sale and use of antibiotics by farmers, and enhance hygienic practices at slaughterhouses.

scholarly journals Use of cefoxitin as indicator for detection of Methicillin Resistant Staphylococcus aureus

2011 ◽  
Vol 8 (4) ◽  
pp. 947-955 ◽  
Author(s):  
Baghdad Science Journal
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Agent ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Critical Diameter ◽  
Brain Heart Infusion ◽  
Diffusion Method ◽  
Disc Diffusion ◽  
Disc Diffusion Method ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant

Rapid and accurate identification of Methicillin Resistant Staphylococcus aureus is essential in limiting the spread of this bacterium. The aim of study is the detection of Methicillin Resistant Staphylococcus aureus (MRSA) and determining their susceptibility to some antimicrobial agent. A total of fifty clinical Staphylococcus aureus, isolated from the nose of health work staff in surgery unit of Kalar general hospital and from ear of patients attended to the same hospital. The susceptibilities of isolates were determined by the disc diffusion method with oxacillin (1 ?g) and cefoxitin (30 ?g), and by the mannitol salt agar supplemented with cefoxitin (MSA-CFOX), susceptibilities of isolates to other antimicrobial agent were determined by standard disc diffusion method, Brain heart infusion (BHI) agar with vancomycin was used for detection of vancomycin resistant Staphylococcus aureus. out of fifty clinical isolates of Staphylococcus aureus 36/50(72%) considered to be MRSA according to MSA-CFOX growth and cefoxitin disc susceptibility results with critical diameter

scholarly journals Panton Valentine Gene in both Community-acquired and Healthcare-acquired Methicillin Resistant Staphylococcus aureus Isolates from Ain Shams University Hospitals

QJM ◽  
2020 ◽  
Vol 113 (Supplement_1) ◽  
Author(s):  
R A Mohamed ◽  
L F Fathi ◽  
N N Salaheldeen
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Cross Sectional Study ◽  
P Value ◽  
University Hospitals ◽  
Cross Sectional ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen that is associated with both hospital and community infections. Panton Valentine leukocidin (PVL) is an important virulence factor of S. aureus that is considered by many authors a marker of community acquired MRSA (CA-MRSA). Aim of the Work This study aimed to determine the prevalence of PVL genes among healthcare acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) and CA-MRSA isolates, and to test the hypothesis that PVL is a reliable marker of CA-MRSA isolates. Material and Methods This comparative cross-sectional study was done on fifteen community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) and fifteen hospital acquired methicillin resistant Staphylococcus aureus (HA-MRSA), obtained from patients attending outpatient clinics, presenting with community-acquired pyogenic infections and patients with healthcare acquired pyogenic infections in Intensive Care Units (ICUs), during the period from May 2017 till February 2018. Clinical specimens included pus and different body fluids. Staphylococcus aureus was isolated and identified using conventional microbiological methods3. Isolates were then tested for methicillin resistance by culture on mannitol salt agar (MSA) with cefoxitin4. The presence of mecA and pvl genes in all MRSA isolates was subsequently detected by PCR5,6. Results Among 15 HA-MRSA isolates, mecA gene was positive in 40% (6/15) of isolates, while pvl gene was positive in 53.3% (8/15) of isolates. Among 15 CA-MRSA isolates, mecA gene was positive in 46.7% (7/15) of isolates, while pvl gene was positive in 26.7% (4/15) of isolates. Conclusion We conclude that pvl gene is not a sole genetic marker for diagnosis CA-MRSA, as there was no significant correlation between mecA that encodes for methicillin resistance and pvl genes among fifteen CA-MRSA isolates (P value =1).

scholarly journals Evaluation of MRSA Chrome agar for the detection of methicillin resistant Staphylococcus aureus

2014 ◽  
Vol 7 (1) ◽  
pp. 1-4
Author(s):  
Durdana Chowdhury ◽  
Sanya Tahmina Jhora ◽  
Tarek Mahbub Khan ◽  
Sadia Afroz
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Clinical Samples ◽  
Clinical Microbiology Laboratory ◽  
Blue Color ◽  
Disc Diffusion ◽  
Diffusion Test ◽  
Gold Standard Method ◽  
Methicillin Resistant

The aim of this study was to evaluate the efficacy of MRSA Chrome agar to detect methicillin resistant Staphylococcus aureus (MRSA) and compare it with 1?g oxacillin disc diffusion tests and detection of mecA gene by PCR. A total 116 Staphylococcus aureus (S. aureus), isolated from various clinical samples, were obtained from three tertiary care hospitals of Dhaka city. S. aureus was identified by colony characters, Gram stain and standard biochemical procedures. MRSA was detected by susceptibility to 1?g oxacillin disc, growth of denim blue color colonies of S. aureus on the Brilliance MRSA Chrome agar at 24 and 48 hours of incubation. PCR was performed for amplification of mecA gene as a gold standard method. Out of 116 isolated S. aureus, 33 (28.44%) were MRSA by oxacillin disc diffusion test where mecA gene was detected in 28 strains. On MRSA Chrome agar, 29 (25.0%) S. aureus produced denim blue colonies at 24 hours, of which 28 isolates possessed mecA gene. At 48 hours incubation, an additional 4 isolates yielded denim blue colonies from which mecA gene could not be identified. All the strains of S. aureus that produced denim blue colonies at 24 and 48 hours were resistant to oxacillin. The sensitivity, specificity and accuracy of oxacillin disc diffusion test were 100%, 94.31% and 95.68% and Chrome agar at 24 hours were 100%, 98.86% and 99.13% respectively. Thus MRSA Chrome agar could be good choice in clinical microbiology laboratory for rapid and accurate identification of MRSA. DOI: http://dx.doi.org/10.3329/imcj.v7i1.17697 Ibrahim Med. Coll. J. 2013; 7(1): 1-4

scholarly journals Mutation-Based Antibiotic Resistance Mechanism in Methicillin-Resistant Staphylococcus aureus Clinical Isolates

2021 ◽  
Vol 14 (5) ◽  
pp. 420
Author(s):  
Tanveer Ali ◽  
Abdul Basit ◽  
Asad Mustafa Karim ◽  
Jung-Hun Lee ◽  
Jeong-Ho Jeon ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Active Site ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Resistance Mechanism ◽  
Ligand Docking ◽  
Clinical Samples ◽  
Allosteric Site ◽  
Methicillin Resistant

β-Lactam antibiotics target penicillin-binding proteins and inhibit the synthesis of peptidoglycan, a crucial step in cell wall biosynthesis. Staphylococcus aureus acquires resistance against β-lactam antibiotics by producing a penicillin-binding protein 2a (PBP2a), encoded by the mecA gene. PBP2a participates in peptidoglycan biosynthesis and exhibits a poor affinity towards β-lactam antibiotics. The current study was performed to determine the diversity and the role of missense mutations of PBP2a in the antibiotic resistance mechanism. The methicillin-resistant Staphylococcus aureus (MRSA) isolates from clinical samples were identified using phenotypic and genotypic techniques. The highest frequency (60%, 18 out of 30) of MRSA was observed in wound specimens. Sequence variation analysis of the mecA gene showed four amino acid substitutions (i.e., E239K, E239R, G246E, and E447K). The E239R mutation was found to be novel. The protein-ligand docking results showed that the E239R mutation in the allosteric site of PBP2a induces conformational changes in the active site and, thus, hinders its interaction with cefoxitin. Therefore, the present report indicates that mutation in the allosteric site of PBP2a provides a more closed active site conformation than wide-type PBP2a and then causes the high-level resistance to cefoxitin.

scholarly journals Detection of Methicillin Resistant Staphylococcus aureus in Dairy Products and Anterior Nares of Dairy Workers

2019 ◽  
Vol 6 ◽  
pp. 59-62
Author(s):  
Ranjana K.C. ◽  
Ganga Timilsina ◽  
Anjana Singh ◽  
Supriya Sharma
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Susceptibility ◽  
Dairy Products ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Dairy Product ◽  
Control Measures ◽  
Diffusion Method ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Dairy Workers

Objectives: To isolate methicillin resistant Staphylococcus aureus (MRSA) from anterior nares of dairy workers and dairy products and assess the antibiotic susceptibility pattern of the isolates. Methods: Swab samples collected from anterior nares of dairy workers and dairy product (butter) were inoculated into mannitol salt agar and incubated at 37ºC for 24 hours. Identification was done based on colony characteristics, Gram's staining, catalase, oxidase and coagulase test. Antibiotic susceptibility testing was done by modified Kirby Bauer disc diffusion method. MRSA was confirmed by using cefoxitin disc. Results: A total of 109 S. aureus (98 from dairy workers and 11 from butter samples) were isolated. Out of them 32 MRSA were isolated from dairy workers and 4 from butter samples. The association between age group and MRSA was found insignificant (p = 0.115). The association of MRSA between male and female workers was found significant (>0.05). About 86% of the MRSA isolates were susceptible to Gentamicin (86.11%) followed by Ciprofloxacin (77.78%). Conclusion: Detection of MRSA among dairy workers and dairy products warrants proper handling and adequate control measures to prevent transmission of MRSA from dairy industry.

scholarly journals Prevalence of Methicillin-Resistant Staphylococcus aureus on Paper Currency Notes

2021 ◽  
pp. 25-32
Author(s):  
Rikhi Ram Marasini ◽  
Pratikshya Shrestha ◽  
Prabhat Dhakal ◽  
Sukra Raj Shrestha ◽  
Sirjana Adhikari ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Brain Heart Infusion ◽  
Biochemical Tests ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Total Prevalence ◽  
Paper Currency ◽  
Antibiotic Susceptibility Test ◽  
Hygiene Measures

The main objective of this study was to determine the prevalence of Methicillin Resistant Staphylococcus aureus (MRSA) in paper currency. The paper currencies in circulation in Pokhara Metropolitan City were inspected. Bills of various denominations (Rs 5, 10, 20, 50, 100, 500 and 1000) were collected from five different locations; namely Food and Vegetable Shop, Bus conductor, Hospital Pharmacy, Butcher Shop and Grocery Shop. Collected sample were cultured and incubated for 24 hours at 37 oC in Brain Heart Infusion (BHI) Broth. The inoculums were further cultured on Mannitol Salt Agar (MSA) and Blood Agar (BA) media to obtain colonies, which were examined and evaluated for various parameters like gram staining and biochemical tests for identification. Then, antibiotic susceptibility test of the isolates was performed using standard procedures. A total of 35 sample of paper currency were processed, all of which showed positive growth. Out of 86 total isolates, 21 (24.42%) were Staphylococcus aureus followed by Coagulase Negative Staphylococci 19 (22.09%), Diptheroids 14 (16.3%), Bacillus spp 13 (15.11%), Micrococci 9 (10.46%), Streptococcus pneumonia 4 (4.65%), Viridans Streptococcus 4 (4.65%) and Streptococcus pyogenes 2 (2.32%). The total prevalence of MRSA in this study was 7 (33.33%). Paper currency contaminated with MRSA poses a high threat to those handling the bills as well as the community. Thus, this study suggests proper hygiene measures to be adopted after handling of paper currency to minimize the risk of contamination and emergence of diseases.

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.

Evaluation of antibiotic resistance pattern and mecA gene frequency in methicillin-resistant Staphylococcus aureus strains collected from clinical specimens in Marand hospital and treatment centers, Iran

2021 ◽  
Author(s):  
Abolfazl Jafari-Sales ◽  
Zahra Sadeghi Deylamdeh ◽  
Afsoon Shariat
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Medical Centers ◽  
Wide Range

Introduction: Staphylococcus aureus causes a wide range of infections and as a multivalent pathogen is one of the causative agents of nosocomial and community infections. Therefore, the aim of this study was to identify and determine the pattern of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in hospitals and medical centers in Marand city and also to evaluate the presence of mecA gene. Materials and Methods: In this cross-sectional descriptive study, 385 samples of S. aureus were collected from different clinical samples of patients in hospitals and medical centers of Marand city. S. aureus was identified using standard biochemical methods.  Methicillin resistance was determined by disk diffusion method in the presence of oxacillin and cefoxitin. The pattern of antibiotic resistance of the strains was determined by disk diffusion method and according to CLSI recommendation and also PCR method was used to evaluate the frequency of MecA gene. Results: In the present study, out of 385 samples of S. aureus, 215 (55.84%) samples were methicillin resistant. PCR results for mecA gene showed that 110 samples had mecA gene.  The highest antibiotic resistance was observed against penicillin (100%) and erythromycin (83.63%). Most MRSA were isolated from urine and wound samples. Conclusion: The results of this study indicate the prevalence of methicillin-resistant species and also the increase in antibiotic resistance of MRSA to various antibiotics.  Therefore, in order to prevent increased resistance to other antibiotics, it is recommended to avoid inappropriate use of antibiotics.

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