Abstract
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.
Highlights
Staphylococcus aureus is usually found to be tightly associated with humans; it can be detected as well in numerous parts of our environment, which include water, dust, feces, air, and clothing [1]
They are producing various factors of virulence, which include toxic shock syndrome toxin (TSST) and staphylococcal enterotoxins (SEs), which are responsible for the staphylococcal food poisoning [9, 10]
A total of 43 isolates of S. aureus (MRSA) and three reference strains were included in evaluating the specificity of PCR and Loopmediated Isothermal Amplification Method (LAMP) assays for the detection of the femA and mecA genes of S. aureus
Summary
Staphylococcus aureus is usually found to be tightly associated with humans; it can be detected as well in numerous parts of our environment, which include water, dust, feces, air, and clothing [1]. S. aureus that includes MRSA is one of the significant bacterial pathogens related to the health-care and community. They are producing various factors of virulence, which include toxic shock syndrome toxin (TSST) and staphylococcal enterotoxins (SEs), which are responsible for the staphylococcal food poisoning [9, 10]. LAMP, that is a procedure based upon the auto-cycling strand displacement DNA synthesis with the use of Bst DNA polymerase enzyme, has been developed in the year 2000 [14] Such Bst DNA polymerase was obtained from the Bacillus stearo-thermophilus, possessing 5ʹ—3ʹ exonuclease activities and requiring high magnesium concentration for the optimal activities [14].
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