scholarly journals Molecular investigation of methicillin-resistant staphylococcus aureus isolates from blood: USA600 emerges as the major type

2018 ◽  
Vol 12 (05) ◽  
pp. 336-341 ◽  
Author(s):  
Mehdi Goudarzi ◽  
Fattaneh Sabzehali ◽  
Mohsen Heidary ◽  
Hadi Azimi ◽  
Hossein Goudarzi
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Diffusion Method ◽  
Major Type ◽  
Molecular Characteristics ◽  
Disk Diffusion Method ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Encoding Genes

Introduction: The widespread emergence of methicillin-resistant Staphylococcus aureus is turning into a real worry in public health. The goals of the present study were to identify resistance and virulence encoding genes and molecular characteristics of methicillin-resistant S. aureus bloodstream isolates. Methodology: A cross-sectional study was conducted on 84 S. aureus bloodstream isolates during a 10-month period. To evaluate antibiotic susceptibility of the isolates, we used Kirby-Bauer disk diffusion method. In addition, the prevalence of antimicrobial resistance and toxins genes was assessed using polymerase chain reaction. Isolates were typed according to polymorphisms seven housekeeping genes by MLST. Results: All the isolates were resistant to methicillin. The most prevalent resistance gene was mecA gene (100%) followed by tetM (57.1%), aac (6΄)-Ie/aph (2˝) (53.6%), ant (4΄)-Ia (46.4%), ermA (45.2%), msrA (35.7%), msrB (33.3%), aph (3΄)-IIIa (33.3%), ermB (31%), ermC (16.7%), and mupA (14.3%) genes. The presence of toxin encoding genes tst, pvl, eta, and etb were detected in 25%, 14.3%, 3.6% and 3.6%, respectively. The isolates were classified into five different sequence types: ST45 (29.8%), ST239 (27.4%), ST858 (21.4%), ST22 (17.8%), and ST59 (3.6%). All the high-level mupirocin-resistant (HLMUPR) strains belonged to ST239, while the low-level mupirocin resistant (LLMUPR) strains belonged to ST22 (13%) and ST239 (6%). Conclusions: To the best of our knowledge, the present study is the first report of ST59 in MRSA bloodstream isolates in Iran. Our data demonstrated the need for thorough epidemiological monitoring to detect emergence and dissemination of MDR- MRSA types in our hospitals.

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.

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 Phenotypic and Genotypic Characteristics of Methicillin-Resistant Staphylococcus aureus Isolated from Dairy and Meat Products in Iran

2020 ◽  
Author(s):  
M.M. Soltan Dallal ◽  
Z. Salehipour ◽  
M.K. Sharifi Yazdi ◽  
R. Bakhtiari ◽  
M. Abdi
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Meat Products ◽  
Gastrointestinal Diseases ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Genotypic Characteristics

Background: Methicillin-Resistant Staphylococcus aureus (MRSA) plays an important role in gastrointestinal diseases. The goal of this research was to determine phenotypic and genotypic characteristics of MRSA isolated from dairy and meat products in Iran. Methods: Ninety-three S. aureus isolates were prepared which had been obtained in our previous study. Antimicrobial susceptibility testing was done using disk diffusion method. The isolates were further analyzed by mecA gene detection. Staphylococcal Enterotoxins (SEs) and Toxic Shock Syndrome Toxin 1 (TSST1) were screened. Biotyping and molecular typing were done by short sequence repeats of spa and coa genes. Results: Five out of 93 S. aureus isolates (5.37%) included mecA. All five MRSA isolates were sensitive to at least six tested antibiotics and none were resistant to vancomycin. Furthermore, two isolates were multidrug resistant. Four isolates produced SEs and TSST1. Three out of 5 isolates were related to human biotype and two belonged to non-host-specific biotype. Conclusion: Presence of MRSA in dairy and meat products may be an important hygienic risk for the Iranian consumers, especially for immunocompromised people.

scholarly journals Occurrence and characteristics of methicillin resistant Staphylococcus aureus and methicillin resistant coagulase-negative staphylococci in raw milk manufacturing

2012 ◽  
Vol 29 (Special Issue) ◽  
pp. S11-S16 ◽  
Author(s):  
M. Vyletělová ◽  
H. Vlková ◽  
I. Manga
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Coagulase Negative Staphylococci ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Staphylococcal Cassette Chromosome

For monitoring the occurrence of MRSA (methicillin resistant Staphylococcus aureus) and MR-CNS (methicillin resistant coagulase-negative staphylococci), cow’s, goat’s, and sheep’s milks (bulk milks and individual samples) were investigated. Human nasal and throat swabs of the farm staff and nasal swabs of animals were also investigated as well. In total 1729 samples were examined and 634 strains were isolated by means of the cultivation method and used in this study. Generic identification of the staphylococci isolates was done performed by biochemical tests and all S. aureus and CNS isolates were checked by the PCR method for the presence of mecA gene which is responsible for methicillin resistance. The presence of the staphylococcal cassette chromosome mec (SCCmec), Panton-Valentine leukocidin (pvl) and genes encoding toxic shock syndrome toxin (tst) was detected in all strains confirmed as MRSA. The species were also examined for antimicrobial susceptibility by using disk diffusion method with antibiotic disks. S. aureus was the most frequently identified species from the samples tested (n = 557; 32.2%), followed by S. haemolyticus (n = 32; 1.9%), S. chromogenes (n = 24; 1.4%), S. epidermidis (n = 20; 1.2%), and S. caprae (n = 1; 0.16%). Among the resistant staphylococci (n = 49), S. aureus (n = 25; 51%) was found the most frequently, followed by S. epidermidis (n = 17; 34.7%), S. chromogenes (n = 6; 12.2%), and S. haemolyticus (n = 1; 2%). The resistant Staphyloccocus sp. occurred mainly in cow’s milk (MRSA, S. epidermidis, S. chromogenes, S. haemolyticus) and in animal’s swabs (S. epidermidis). One MRSA was also found in goat’s milk and one was isolated from human swab. No resistant strains were found in sheep’s milk. The negative results of the analysed genes presence (pvl, tst) were identical with all MRSA tested. The staphylococcal cassette chromosome mec (SCCmec) was classified as type IV or V.

scholarly journals Predominance of PVL-negative community-associated methicillin-resistant Staphylococcus aureus sequence type 8 in newly diagnosed HIV-infected adults, Tanzania

2021 ◽  
Author(s):  
Joel Manyahi ◽  
Sabrina J. Moyo ◽  
Said Aboud ◽  
Nina Langeland ◽  
Bjørn Blomberg
Keyword(s):  
Staphylococcus Aureus ◽  
Hiv Infection ◽  
Diffusion Method ◽  
Molecular Characteristics ◽  
People Living With Hiv ◽  
Newly Diagnosed ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Inducible Clindamycin Resistance

AbstractDifficult-to-treat infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are of concern in people living with HIV infection as they are more vulnerable to infection. We aimed to identify molecular characteristics of MRSA colonizing newly diagnosed HIV-infected adults in Tanzania. Individuals newly diagnosed with HIV infection were recruited in Dar es Salaam, Tanzania, from April 2017 to May 2018, as part of the randomized clinical trial CoTrimResist (ClinicalTrials.gov identifier: NCT03087890). Nasal/nasopharyngeal isolates of Staphylococcus aureus were susceptibility tested by disk diffusion method, and cefoxitin-resistant isolates were characterized by short-reads whole genome sequencing. Four percent (22/537) of patients carried MRSA in the nose/nasopharynx. MRSA isolates were frequently resistant towards gentamicin (95%), ciprofloxacin (91%), and erythromycin (82%) but less often towards trimethoprim-sulfamethoxazole (9%). Seventy-three percent had inducible clindamycin resistance. Erythromycin-resistant isolates harbored ermC (15/18) and LmrS (3/18) resistance genes. Ciprofloxacin resistance was mediated by mutations of the quinolone resistance-determining region (QRDR) sequence in the gyrA (S84L) and parC (S80Y) genes. All isolates belonged to the CC8 and ST8-SCCmecIV MRSA clone. Ninety-five percent of the MRSA isolates were spa-type t1476, and one exhibited spa-type t064. All isolates were negative for Panton-Valentine leucocidin (PVL) and arginine catabolic mobile element (ACME) type 1. All ST8-SCCmecIV-spa-t1476 MRSA clones from Tanzania were unrelated to the globally successful USA300 clone. Carriage of ST8 MRSA (non-USA300) was common among newly diagnosed HIV-infected adults in Tanzania. Frequent co-resistance to non-beta lactam antibiotics limits therapeutic options when infection occurs.

scholarly journals Methicillin-resistant Staphylococcus aureus bloodstream infection: risk factors and clinical outcome in non-intensive-care units

2012 ◽  
Vol 45 (2) ◽  
pp. 189-193 ◽  
Author(s):  
Karinne Spirandelli Carvalho Naves ◽  
Natália Vaz da Trindade ◽  
Paulo Pinto Gontijo Filho
Keyword(s):  
Risk Factors ◽  
Staphylococcus Aureus ◽  
Bloodstream Infection ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Staphylococcus Aureus Bloodstream Infection

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is spread out in hospitals across different regions of the world and is regarded as the major agent of nosocomial infections, causing infections such as skin and soft tissue pneumonia and sepsis. The aim of this study was to identify risk factors for methicillin-resistance in Staphylococcus aureus bloodstream infection (BSI) and the predictive factors for death. METHODS: A retrospective cohort of fifty-one patients presenting bacteraemia due to S. aureus between September 2006 and September 2008 was analysed. Staphylococcu aureus samples were obtained from blood cultures performed by clinical hospital microbiology laboratory from the Uberlândia Federal University. Methicillinresistance was determined by growth on oxacillin screen agar and antimicrobial susceptibility by means of the disk diffusion method. RESULTS: We found similar numbers of MRSA (56.8%) and methicillin-susceptible Staphylococcus aureus (MSSA) (43.2%) infections, and the overall hospital mortality ratio was 47%, predominantly in MRSA group (70.8% vs. 29.2%) (p=0.05). Age (p=0.02) was significantly higher in MRSA patients as also was the use of central venous catheter (p=0.02). The use of two or more antimicrobial agents (p=0.03) and the length of hospital stay prior to bacteraemia superior to seven days (p=0.006) were associated with mortality. High odds ratio value was observed in cardiopathy as comorbidity. CONCLUSIONS: Despite several risk factors associated with MRSA and MSSA infection, the use of two or more antimicrobial agents was the unique independent variable associated with mortality.

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.

Assessment of Mupirocin Resistance among Clinical Isolates of Methicillin Resistant Staphylococcus aureus

2021 ◽  
Vol 30 (1) ◽  
pp. 109-114
Author(s):  
Nancy M. Attia ◽  
Abeer Abd El Rahim Ghazal ◽  
Omnia M. Khaleel ◽  
Ahmed Gaballah
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Mupirocin Resistance ◽  
High Level ◽  
Research Institute Hospital ◽  
Low Prevalence ◽  
Mrsa Colonization

Background: Methicillin-resistant Staphylococcus aureus (MRSA) colonization is considered a major risk factor for nosocomial infections and its decolonization has reduced these infections. Mupirocin (MUP) is the topical antibiotic of choice for decolonization. MUP decolonization failure is attributed to MUP resistance. Objective: The aim of the current study is to assess MUP resistance among MRSA isolates phenotypically and genotypically. Methodology: Fifty MRSA isolates were identified in Microbiology Department in the Medical Research Institute hospital, Alexandria University. Antibiotic susceptibility to different classes of antibiotics by disk diffusion method was done. MUP minimum inhibitory concentration (MIC) was determined phenotypically by MUP Ezy MIC™ Strips. MUP resistance was determined genetically by multiplex PCR detection of mupA and mupB. Results: Of all MRSA isolates, 6% exhibited high level and none showed low level MUP resistance. Only mupA was detected in all resistant isolates. Conclusion: Despite low prevalence of MUP resistance, it is appropriate to test MUP resistance prior nasal decolonization

scholarly journals Genetic Characterization of Methicillin-Resistant Staphylococcus aureus Isolates from Human Bloodstream Infections: Detection of MLSB Resistance

Antibiotics ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 375
Author(s):  
Vanessa Silva ◽  
Sara Hermenegildo ◽  
Catarina Ferreira ◽  
Célia M. Manaia ◽  
Rosa Capita ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bloodstream Infections ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Accessory Gene ◽  
Spa Typing ◽  
Disk Diffusion Method ◽  
Methicillin Resistant

In this study we aimed to characterize antimicrobial resistance in methicillin-resistant Staphylococcus aureus (MRSA) isolated from bloodstream infections as well as the associated genetic lineages of the isolates. Sixteen MRSA isolates were recovered from bacteremia samples from inpatients between 2016 and 2019. The antimicrobial susceptibility of these isolates was tested by the Kirby–Bauer disk diffusion method against 14 antimicrobial agents. To determine the macrolide–lincosamide–streptogramin B (MLSB) resistance phenotype of the isolates, erythromycin-resistant isolates were assessed by double-disk diffusion (D-test). The resistance and virulence genes were screened by polymerase chain reaction (PCR). All isolates were characterized by multilocus sequence typing (MLST), spa typing, staphylococcal chromosomal cassette mec (SCCmec) typing, and accessory gene regulator (agr) typing. Isolates showed resistance to cefoxitin, penicillin, ciprofloxacin, erythromycin, fusidic acid, clindamycin, and aminoglycosides, confirmed by the presence of the blaZ, ermA, ermC, mphC, msrA/B, aac(6’)-Ie-aph(2’’)-Ia, and ant(4’)-Ia genes. Three isolates were Panton–Valentine-leukocidin-positive. Most strains (n = 12) presented an inducible MLSB phenotype. The isolates were ascribed to eight spa-types (t747, t002, t020, t1084, t008, t10682, t18526, and t1370) and four MLSTs (ST22, ST5, ST105, and ST8). Overall, most (n = 12) MRSA isolates had a multidrug-resistance profile with inducible MLSB phenotypes and belonged to epidemic MRSA clones.

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