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 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).

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 Findings of methicillin-resistant strains of Staphylococcus aureus in livestock

2010 ◽  
Vol 27 (Special Issue 2) ◽  
pp. 36-41
Author(s):  
Z. Šťástková ◽  
S. Karpíšková ◽  
R. Karpíšková
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Potential Hazard ◽  
The Czech Republic ◽  
Methicillin Resistant ◽  
Milk Samples ◽  
Genes Encoding ◽  
Resistant Strains ◽  
Bulk Tank

The aim of our study was to determine the occurrence of methicillin resistant Staphylococcus aureus (MRSA) at dairy farms in the Czech Republic. Altogether 1061 samples from 95 farms were examined. The samples analysed were milk (individual and bulk tank milk samples), animal swabs and swabs from the farm environment. In total, 299 S. aureus isolates were obtained, of which 23 were MRSA. These MRSA isolates originated from three farms (13 isolates from farm A and 5 isolates from each of farms B and C). All MRSA isolates carried the mecA gene while none of them carried the genes for PVL, TSST-1 and exfoliatins. Only the isolates from goat farm C were positive for the genes encoding enterotoxins. By SCCmec typing, the strains were classified as community-associated MRSA carrying SCCmec IV or V. This study revealed that animals can be an important source of methicillin resistant staphylococci and represent a potential hazard of further spread.

scholarly journals Toxin production and drug resistance profiles of pediatric methicillin-resistant Staphylococcus aureus isolates in Tehran

2017 ◽  
Vol 11 (10) ◽  
pp. 759-765 ◽  
Author(s):  
Mozhgan Esmaeili Benvidi ◽  
Hamidreza Houri ◽  
Zohreh Ghalavand ◽  
Bahram Nikmanesh ◽  
Hadi Azimi ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Toxin Production ◽  
Cross Sectional Study ◽  
Biochemical Tests ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Present Cross Sectional Study ◽  
Invasive Diseases

Introduction: Staphylococcus aureus is known to be a major cause of skin and soft tissue infections, pneumonia and invasive diseases. In this study, attempts were made to examine the prevalence of tsst-1, eta, etb, and luk-PV genes among methicillin-resistant S. aureus (MRSA) isolated from children in Tehran. Methodology: In the present cross-sectional study, a total of 100 MRSA were isolated from children who were referred to a pediatric hospital during 11-month period of September 2014 to August 2015. Isolates were identified using biochemical tests and then, using PCR, the isolates were tested for the presence of mecA, tsst-1, eta, etb, and luk-PV genes. Susceptibility of isolates to cefoxitin, penicillin, erythromycin, clindamycin, gentamicin, rifampin, minocycline, co-trimoxazole, linezolid, and vancomycin were evaluated using standard methods. Results: It was found that the MRSA isolates had the greatest resistance to clindamycin (72%) and erythromycin (59%), while the lowest rates of resistance were observed to be related to minocycline (6%) and rifampin (12%). All of isolates were sensitive to vancomycin and linezolid. The mecA gene was detected in all the isolates. Moreover, luk-PV and tsst-1 were detected in 18% and 17% of the isolates, respectively. None of the isolates harbored eta and etb genes. Conclusions: Our data provide specifications about the toxin production status of S. aureus isolates from pediatric children. The current study showed increased resistance to different antibiotics in S. aureus isolates. Therefore, to prevent multi-resistance to other antibiotic classes, it is essential to withhold prescriptions and stop unessential use of available antibiotics.

scholarly journals Prevalence of Panton-Valentine leukocidin genes in community-associated methicillin-resistant Staphylococcus aureus in the district of Pomoravlje

2016 ◽  
Vol 73 (3) ◽  
pp. 256-260 ◽  
Author(s):  
Ljiljana Petrovic-Jeremic ◽  
Nada Kuljic-Kapulica ◽  
Elizabeta Ristanovic ◽  
Dragana Josic ◽  
Zorica Lepsanovic
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Clinical Laboratory ◽  
Meca Gene ◽  
Latex Agglutination ◽  
The Other ◽  
Methicillin Resistant ◽  
Standard Polymerase Chain Reaction ◽  
Panton Valentine Leukocidin ◽  
Valentine Leukocidin

Background/Aim. Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains appear to have rapidly disseminated among population in the community without established risk factors for MRSA worldwide. Panton-Valentine leukocidin (PVL) is a cytolytic toxin, encoded by the lukF-PV and lukF-PV genes. PVL may be the key toxin responsible for enhanced virulence of CA-MRSA. The aim of this study was to detect the genes encoding PVL in CA-MRSA isolates from healthy people from the District of Pomoravlje. Methods. We took throat and nose swabs from healthy, employed persons with mandatory sanitary examinations and analyzed the presence of MRSA, between January 2011 and December 2012 in the District of Pomoravlje. Susceptibility of isolated strains to cefoxitin was investigated by using disc diffusion according to the recommendation of CLSI (Clinical Laboratory Standard Institute), and by E test. The presence of penicillin-binding protein 2a (PBP2a) in Staphylococci was detected using latex agglutination Slidex ?MRSA Detection test. The gold standard, polymerase chain reaction (PCR) assay, was used for detection of mecA gene and PVL gene, and typing of SCCmec region. Results. Our investigation showed that staphylococcal carrier state was present in 2.58% of 52,910 throat and nasal swabs, and in 50 of them (3.67%) MRSA was isolated. Among these MRSA, 2 (4%) isolates were PVL-positive. Conclusion. The prevalence of CAMRSA and the presence of PVL gene among healthy, employed population in the District of Pomoravlje were low. The values obtained in this study show that, our region is not significantly different from the other parts of our country, nor from the other European countries.

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.

scholarly journals Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Sccmec Type ◽  
Meca Gene ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

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 Methicillin-resistant Staphylococcus aureus on surfaces of an Intensive Care Unit

2011 ◽  
Vol 24 (4) ◽  
pp. 453-458 ◽  
Author(s):  
Adriano Menis Ferreira ◽  
Denise de Andrade ◽  
Marcelo Alessandro Rigotti ◽  
Margarete Teresa Gottardo de Almeida
Keyword(s):  
Intensive Care Unit ◽  
Staphylococcus Aureus ◽  
Intensive Care ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Descriptive Analysis ◽  
Infusion Pump ◽  
Cross Sectional Study ◽  
Cross Sectional ◽  
Methicillin Resistant

OBJECTIVE: To evaluate the presence of methicillin-resistant Staphylococcus aureus (MRSA) in areas close to patients in a General Intensive Care Unit. METHODS: This is a cross-sectional study, in which microbiological samples were collected from five surfaces (left / right bed siderails, bed crank, table, buttons on the infusion pump, and cotton gowns) from each of ten patient rooms, totaling 63 samples. To collect samples, the Petri FilmTM Staph Express Count System 3M TM was used to screen for methicillin resistance, with the Mueller-Hinton agar supplemented with 4% sodium chloride and 6 µg / ml of oxacillin. Descriptive analysis was conducted to determine the frequency (n) and percentage (%) of contamination of environmental surfaces. RESULTS: Of 48 samples positive for Staphylococcus aureus, 29 (60.4%) were resistant to methicillin. The incidence on the siderails and bed cranks, table, buttons on the infusion pumps and aprons were, respectively, 55.5%, 57.1%, 57.1%, 60.0% and 75.0%. CONCLUSION: The results suggest that the surfaces around the patient constitute a major threat, as they represent secondary reservoirs of MRSA.

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