The Molecular Detection of mecA Genes of Staphylococcus Aureus

2019 ◽  
Vol 2 (1) ◽  
Author(s):  
Cynthia N Nwaogaraku
Keyword(s):  
Staphylococcus Aureus ◽  
Pathogenic Bacteria ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Diffusion Method ◽  
Gene Detection ◽  
Blood Samples ◽  
Life Threatening ◽  
Polymerase Chain ◽  
Mrsa Strains

Antibiotic resistance is common among pathogenic bacteria associated with community acquired and nosocomial infections. Methicillin-resistant Staphylococcus aureus (MRSA) infections have become a global health problem particularly in hospital setup causing simple skin infections to life threatening infections. The present study aimed to investigate the presence of mecA genes in MRSA from pigs, using Polymerase Chain Reaction. One hundred S. aureus isolates of blood samples from Pigs in Bariga, Lagos State were collected from Molecular Biology and Biotechnology Unit, Nigeria Institute of Medical Research. Methicillin resistance was determined by Kirby-Bauer’s disc diffusion method. The PCR was used for mecA gene detection from MRSA strains. Twenty-five pure Staphylococcus aureus isolates were identified based on cultural characteristics, biochemical reactions and positive slide coagulase test. Out of these, 11 (44%) strains were MRSA by phenotypic method. Amplification of mecA gene for all the 11 MRSA isolates was negative when visualized on 2% agarose gel electrophoresis. Eleven strains of MRSA were found among Staphylococcus aureus isolates of blood samples from Pigs. The MRSA phenotype observed in the isolates was not the classical mecA mediated resistance. Hence, it is highly recommended to consider alternative mechanisms for β-lactams resistance that may compete with mecA gene in the emergence of MRSA phenomenon in Nigeria.

scholarly journals Bacterial Etiology of Wound Exudates in Tertiary Care Cancer Patients and Antibiogram of the Isolates

2020 ◽  
Vol 13 ◽  
pp. 117863372095207
Author(s):  
Kiran Duwadi ◽  
Sujan Khadka ◽  
Sanjib Adhikari ◽  
Sanjeep Sapkota ◽  
Pabitra Shrestha
Keyword(s):  
Staphylococcus Aureus ◽  
Pathogenic Bacteria ◽  
Methicillin Resistance ◽  
Tertiary Care ◽  
Antibiotic Sensitivity ◽  
Cross Sectional Study ◽  
Diffusion Method ◽  
Disc Diffusion ◽  
Biochemical Tests ◽  
Cross Sectional

Introduction: Patients with malignancies frequently develop infections as a result of surgical procedures and fungating wounds leading to pus formation. This cross-sectional study was conducted to explore the bacteriological spectra of infections of various cancer sites and their antibiotic sensitivity patterns among the patients visiting minor operation theatre (OT) of B.P. Koirala Memorial Cancer Hospital (BPKMCH), Chitwan, Nepal. Methods: Over a period of 3 months from September to November 2018, a total of 183 wound exudates and pus samples were collected and analyzed by standard microbiological procedures. Isolates were identified based on the colony characters, Gram staining and an array of biochemical tests. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion technique according to criteria set by CLSI, 2016. Methicillin resistance in Staphylococcus aureus was tested with the help of cefoxitin using disc diffusion method. Results: Out of the 183 samples, 149 (81.4%) were culture positive. Among 13 different isolates identified, S. aureus (43.0%) was predominant followed by E. coli (14.0%). Higher incidence of bacteria was seen among the males (52.3%), in the age group 51 to 60 years (26.8%) and among the patients undergoing surgical intervention to deal with cancer (34.2%). The prevalence of wound infection was significantly affected by gender, age, and treatment regimen ( P < .01). Out of the total 68 S. aureus isolates, 38 (44.1%) were deemed as Methicillin-resistant Staphylococcus aureus (MRSA). Among the 158 isolates, 85 (53.8%) were multi-drug resistant (MDR). Cefepime was the most effective antibiotic for Gram positive isolates whereas both imipenem and meropenem were found to be equally more effective for Gram negative isolates. Conclusion: This study suggests that patients with malignancies harbor pathogenic bacteria; therefore, prudent use of antibiotics is essential to prevent the emergence of MDR pathogens.

scholarly journals Potensi Ekstrak Atuna racemosa sebagai Anti - Methicillin Resistant Staphylococcus aureus (MRSA)

2017 ◽  
Vol 35 (2) ◽  
pp. 260
Author(s):  
Siti Isrina Oktavia Salasia ◽  
Novra Arya Sandi ◽  
Fajar Budi Lestari ◽  
Verda Farida ◽  
Nurbani Aziz
Keyword(s):  
Staphylococcus Aureus ◽  
Meca Gene ◽  
New Drugs ◽  
Diffusion Test ◽  
Methicillin Resistant ◽  
Causative Agents ◽  
Mrsa Infection ◽  
Polymerase Chain ◽  
Mrsa Strains ◽  
Multiple Antibiotics

Staphylococcus aureus is one of the major causative agents of mastitis in animals and a variety of human diseases such as septicemia, endocarditis, arthritis dan osteomyelitis. Infection of Methicillin-resistant S. aureus (MRSA) has been widely reported and these strains are usually resistant to multiple antibiotics. The purpose of this study was to evaluate the potential of Atuna racemosa, as an alternative herbal medicine against MRSA infection. The MRSA strains were isolated from human and confirmed based on their resistant to various antibiotics and analyzing of the mecA gene by polymerase chain reaction (PCR). Atuna racemosa originated from Ambon, Maluku, Indonesia, were extracted using 70% ethanol. The activities of the Atuna racemosa extract against MRSA were performed by diffusion disc agar and dilussion agar tests. The results showed that Atuna racemosa extract has the barrier effect of MRSA growth at a concentration of 5% in the diffusion test and at a concentration of 7% in the dilution test. Atuna racemosa could be used as an alternative new drugs with dose of 0.07 g/ml (7%) against MRSA which is multi-resistant to many antibiotics. 

scholarly journals Characteristics of Erythromycin Resistance in Methicillin-Resistant Staphylococcus aureus Isolated From Raw Milk

2019 ◽  
Vol 7 (4) ◽  
pp. 121-125
Author(s):  
Fatemeh Mahdavi ◽  
Fatemeh Zaboli ◽  
Rahem Khoshbakht
Keyword(s):  
Staphylococcus Aureus ◽  
Resistance Genes ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Raw Milk ◽  
Erythromycin Resistance ◽  
Methicillin Resistant ◽  
Inducible Clindamycin Resistance ◽  
Mrsa Strains

Background: Methicillin-resistant Staphylococcus aureus (MRSA) strains are one of the most important multidrug resistant microorganisms that threaten human health. Objective: The present study was conducted to evaluate genotypic and phenotypic characteristics of erythromycin resistance among MRSA isolates recovered from raw milk in Iran. Materials and Methods: A total of 50 MRSA isolates were recovered from raw milk. Tests for erythromycin and clindamycin susceptibility and inducible clindamycin resistance were done. In addition, the presence of the methicillin resistance determinant (mecA), erythromycin resistance genes (ermA, ermB, ermC and msrA) and an important virulence gene (Panton– Valentine leukocidin) were investigated using polymerase chain reaction (PCR) method. Results: Forty-eight percent (24/50) and 46% (23/50) of the isolates were resistant to erythromycin and clindamycin, respectively. Seven (14%) isolates showed inducible clindamycin resistance phenotype. The mecA gene was detected in 88% (44/50) of MRSA isolates. The incidence of the ermA, ermB, ermC and msrA genes was 14%, 64%, 12%, and 26%, respectively and the PVL gene was present in 18% (9/50) of MRSA isolates. Conclusion: According to the results of the study, the incidence of erythromycin resistance genes and inducible clindamycin-resistant MRSA strains was high in raw milk samples in Iran.

Molecular Investigation of Staphylococcal Cassette Chromosome mec (SCCmec) Elements Isolated from Intensive Care Unit

2020 ◽  
Author(s):  
Fahimeh Nourbakhsh ◽  
Vajiheh Nourbakhsh ◽  
Samaneh Borooni ◽  
Elaheh Tajbakhsh ◽  
Dana Daneshmand
Keyword(s):  
Intensive Care Unit ◽  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Intensive Care ◽  
Methicillin Resistance ◽  
Diffusion Method ◽  
Clinical Samples ◽  
High Morbidity ◽  
Molecular Investigation ◽  
Mrsa Strains

Background and Aims: Based on the results, Staphylococcus aureus is one of the serious infectious agents found in community and hospitals with remarkable potential for high morbidity and mortality around the globe. The present study was carried out for molecular investigation of methicillinresistant Staphylococcus aureus strains and Staphylococcal Chromosomal Cassette mec (SCCmec) phenotypes isolated from the intensive care unit in Hazrat Fatemeh Zahra hospital of Isfahan. Materials and Methods: A total of 76 clinical wound samples were collected from Hazrat Fatemeh Zahra Hospital in Isfahan and evaluated by polymerase chain reaction (PCR) methods. The Methicillin resistance Staphylococcus aureus (MRSA) screening was performed by genotypic and phenotypic methods; also antibiotic resistance pattern was determined by using the disk diffusion method and related genes by PCR. Results: Totally, 53 (69.7%) out of 76 clinical samples were positive for MRSA. Of the 76 MRSA strains, 39 (63.51%) were PVL positive (51.3%). The most commonly infected samples were collected from wounds (40.8%). The most commonly detected antibiotic resistance genes were mecA (89.61%), tetK (88.23%), tetM (49.15%) and msrA (46.93%). Resultantly, it was shown that MRSA has the highest level of resistance against methicillin (98%), penicillin (97.24%), tetracycline (89.64%). It was also revealed that the most commonly detected SCCmec types in the MRSA strains are types II (14.53%) and III (16.82%). Conclusions: In summary, this paper argues that the orderly surveillance of hospital-associated infections and initial management and supervision of the antibiotic resistance patterns are required to control the prevalence of MRSA.

scholarly journals Prevalence of Methicillin Resistant Staphylococcus Aureus (MRSA) In A Secondary Care Hospital In North Eastern Part of India

2018 ◽  
Vol 2 (1) ◽  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Antibiotic Sensitivity ◽  
Diffusion Method ◽  
Care Hospital ◽  
Wound Infections ◽  
Methicillin Resistant ◽  
Sensitivity Pattern ◽  
Mrsa Strains

Aim: In today’s scenario, Nosocomial infection is a foremost problem in world. Methicillin resistant staphylococcus aureus (MRSA) strains are not only resistant to several antibiotics but also, show a particular ability to spread in hospitals of many countries. The main objective of the present study is to determine the prevalence of MRSA in surgical wound infections and also to define the antimicrobial susceptibility patterns of the strains isolated. Materials and Methods: A total of 133 culture positive Staph.aureus were taken from surgical site wound infections for the study during the period from March 2016 to February 2017. Routine Antibiotic Susceptibility testing was performed and interpreted as per latest CLSI guidelines. Methicillin resistance was detected using cefoxitin disc diffusion method. Results: In total of 133 samples, 81 (60.9%) were found to be MRSA. However, no strain was resistant to Vancomycin, Linezolid or Teichoplanin. Conclusion: Vigorous antimicrobial stewardship and strengthened infection control practices are mandatory to prevent spread and reduce emergence of resistance. Customary surveillance of hospital-associated infection and monitoring of antibiotic sensitivity pattern is required to reduce MRSA prevalence.

scholarly journals Multiplex Polymerase Chain Reaction Based Typing of Staphylococcal Chromosomal Cassette mec of Methicillin Resistant Staphylococcus aureus from Selected Hospitals in Lahore

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Romeeza T ◽  
Naeem R ◽  
Nadeem A
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Sccmec Type ◽  
Meca Gene ◽  
Methicillin Resistant ◽  
Different Types ◽  
Type Ia ◽  
Polymerase Chain ◽  
Sccmec Types

Introduction: Staphylococcus aureus is one of the leading causes of nosocomial infections. Prevalence of methicillin resistant Staphylococcus aureus (MRSA) has been reported in up to 42% of isolates in Pakistan. Methicillin resistance is encoded by the mecA gene and it is transferred by a unique genetic vector called staphylococcal chromosomal cassette (SCCmec). Depending upon the selection pressures of antibiotics, different types of SCCmec elements may prevail in different parts of the world. Materials and Methods: Therefore, this study was designed to fi nd out the major SCCmec types present in selected hospitals of Lahore. For this purpose thirty fi ve MRSA isolates were collected. These strains were reconfi rmed and SCCmec types were determined by multiplex PCR. Results: It was found that 21 (60%) isolates possess SCCmec type IA while 14 (40%) isolates possess SCCmec type IIC. Conclusion: These cassettes are shown to be multiresistant and have not been reported in other Asian countries so far.

EVALUATION OF THE METHOSD FOR DETECTION METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS.

2013 ◽  
pp. 25-31
Author(s):  
Thi Kim Chi Nguyen ◽  
Dinh Binh Tran ◽  
Thi Nam Lien Nguyen ◽  
Van Tuan Mai ◽  
Godreuil Sylvain
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Antibiotic Sensitivity ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Antibiotic Resistant ◽  
Methicillin Resistant ◽  
Resistant Strains

Objective: To evaluate the infections that caused by Methicillin-resistant Staphylococcus aureus and the value of the tests to detect Methicillin-resistant Staphylococcus aureus. Subjects and Methods: Used routine techniques to culture and isolate S.aureus, test the antibiotic sensitivity by Kirby-Bauerr, determination the Methicillin-resistant Staphylococcus aureus by Oxacillin and cefoxitin disc and PCR in identified the mecA gene Staphylococcus aureus. Results: The rate of Staphylococcus aureus isolated is highest which isolated from pus specimens (55.06%). In 267 strains of Staphylococcus aureus isolated in the Department of Microbiology, Hue Central Hospital the Methicillin resistance Staphylococcus aureus was 61.42%. The level of antibiotic resistant strains of Methicillin-resistant Staphylococcus aureus is higher than that in Methicillin-sensitive strains. Conclusion: Cefoxitin 30 microg disk diffusion method to detect Methicillin resistance is effective for determinate Methicillin-resistant Staphylococcus aureus (sensitivity and specificity are all 100.00%). Key words: Staphylococcus aureus Methicillin-resistant.

scholarly journals Detection of Methicillin Resistance in Staphylococcus Aureus by Polymerase Chain Reaction and Conventional Methods: A Comparative Study

2012 ◽  
Vol 4 (02) ◽  
pp. 083-088 ◽  
Author(s):  
Manju M Pillai ◽  
Ragunathan Latha ◽  
Gautam Sarkar
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Chain Reaction ◽  
Mannitol Salt Agar ◽  
Polymerase Chain ◽  
Conventional Methods ◽  
Phenotypic Methods

ABSTRACT Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen worldwide, which has emerged over the past 30 years as a leading cause of both nosocomial and community-acquired infections. Accurate and rapid identification of MRSA in clinical specimens is essential for timely decision on effective antimicrobial chemotherapy. Aim: The present study was conducted to compare two conventional phenotypic methods, oxacillin disk diffusion (ODD) Aim: The present study was conducted to compare two conventional phenotypic methods, oxacillin disk diffusion (ODD) method and mannitol salt agar (MSA) with oxacillin, with polymerase chain reaction (PCR) for mecA gene (as standard). Materials and Methods: A total of 165 consecutive clinical isolates of S. aureus received at the Department of Microbiology in our tertiary care teaching hospital were included in the study. All the isolates were subjected to ODD (1 μg) method, culture in MSA with oxacillin, and PCR for mecA gene. Results: The sensitivity and specificity of ODD test were found to be 93.5% (86.4-97.3%) and 83.5% (79.2-85.8%), respectively, and that of MSA with oxacillin were found to be 87.1% (79.5-92.3%) and 89.3% (84.8-92.5%), respectively. The time taken for diagnosing MRSA by conventional methods is 48-72 h, which is more as compared to PCR which takes 18-24 h. Conclusion: This study recommends advocating PCR for mecA gene on a regular basis for detecting methicillin resistance in S. aureus isolates isolated from sterile body fluids or from special units such as intensive care units.

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.

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