Staphylococcus aureus, which includes the methicillin-resistant S. aureus (MRSA), is a significant human pathogen producing different toxins and results in many different infection types, which include bacteremia, soft-tissue infections, as well as staphylococcal food poisoning. S. aureus is an important food-borne pathogen of humans due to ingestion of food containing enterotoxigenic strains. Detecting S. aureus femA and mecA genes was evaluated with the use of a Loop-mediated Isothermal Amplification Method (LAMP). The accuracy of this approach was similar to that attained using the approach of the conventional polymerase chain (PCR). Those two methods characterized 43 isolates of MRSA which were separated from different samples of foods and were not detected in the two other non-Staphylococcus strains (standard strains). The optimal temperature for the LAMP assay was 65°C, with a detection limit of 2.5ng/μL and 103cfu/ml, when compared to 12.5 ng/μL and 104cfu/ml for PCR. The LAMP assay permits a one-step characterization of a specific gene, with no special equipment, and needs less time compared to the traditional PCR. It is assumed that the LAMP assay is a promising alternative method for the rapid identification of S. aureus and could be used in resource-limited laboratories and fields.
Foodborne diseases are a major risk for human health. Millions of people become sick as a result of eating contaminated food with microorganisms that cause diseases. S. aureus is considered as one of the most important pathogenic bacteria, having the ability to activate certain genes that encode for heat stable enterotoxins and cause Staphylococcal food poisoning. Thus, this study aimed to determine the prevalence of multi resistant Staphylococcus aureus that produce enterotoxins in different sources of food . Forty nine isolates were identified as S.aureus, according to morphological and biochemical tests. They were isolated from 387 different food samples from several randomly covered restaurants and supermarkets in different regions of Baghdad. Molecular diagnosis of S. aureus using specific primers for the 16S rRNA gene was carried out by Polymerase Chain Reaction (PCR ) technique . Susceptibility of 43 isolates of S.aureus was tested against 15 antimicrobial agents. The results revealed that all the isolates were resistant (100%) to mecillinam, highly resistant to vancomycine and meropenemin (74.4 %) and moderately resistant to Oxacillin, Erythromycin Cefotaxime, and Cefiximein (67.4, 60.4, 62.8, , 60.5 %, respectively), while they showed low resistance to Gentamicin (34.8%). In addition, all of these isolates were susceptible to Tigecycline and Amoxicillin/ clavulanic acid and Cefoxitin-Cloxacilin. High percentages of oxicillin resistant S. aureus were isolated from cooked food samples, followed by meat products, and with less percentage from pastry products. Molecular detection of enterotoxins A and B of Staphylococcus aureus isolates was performed using specific primers based on PCR. The results revealed that S. aureus isolated from cooked food had the highest percentage of the isolates producing the enterotoxins A and B. Type A enterotoxin gene showed a higher prevalence than type B gene among cooked food , dairy products and pastry. In conclusion, the results revealed a high prevalence of some classical enterotoxin genes in multi-drug resistant S.aureus isolated from different sources of food, which can cause food-poisoning and, consequently, a potential serious problem for public health.
There is so little information available in Iraq about genetic variability in methicillin resistant Staphylococcus aureus (MRSA), so current study aimed to use six tandem repeat loci of multilocus variable number tandem repeat (MLVA) typing to discriminate among MRSA, so to achieve the aim of this study, six loci, clfA, clfB, sdrC, spa, sspa and sav1078 were selected for multiplex PCR. The PCR product were subjected to capillary electrophoresis by using ABI-Genetic analyzer, then the data were analyzed by using GeneMapper™ Software 5. Fragment sizes were converted into repeats number. The total number of repeats are used to generate allelic profile. The allelic profile used to draw the minimum spanning tree and dendrogram , all the 85 MRSA isolates are clustered into 54 MLVA type.
The study was aimed to evaluate the prevalence of MRSA in some Iraqi hospitals and determine the most powerful methods for identification of MRSA, in order to achieve the, 278 samples were collected from different hospitals in Iraq in various intervals, 204 out of 287 were identified as Staphylococcus aureus by conventional cultural methods and microscopic characteristics and 177 isolates are identified as MRSA by using HiCrome MeReSa Agar Base medium, but 154 of 177 (87%) isolates are methicillin resistance in sensitivity test. MRSA isolates were highly resistant to β-lactam antibiotics and considered multidrug resistant (MDR) in percent of (94.9%). Touchdown PCR used to identify the isolates, 97.05% were identified as Staphylococcus aureus, while 80.88% as MRSA.
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