MOLECULAR DETECTION OF STAPHYLOCOCCUS AUREUS ENTEROTOXINS B USING LOOP-MEDIATED ISOTHERMAL AMPLIFICATION

Background and Aim: Enterotoxins B of Staphylococcus aureus (SEB) is virulence factor in poisonous of this bacteria that highly resistant against heat and pasteurization. Therefore, rapid detection of SEB is very important. The goal of this work is molecular detection of SEB in genome of S. aureus using loop-mediated isothermal amplification (LAMP). Methods: In this study, the Primer explorer v4 software was used for designing specific primers. After culturing of S. aureus, DNA genomic was extracted. LAMP reaction contains Bst buffer, dNTP, Betaine, FIP and BIP ( inner primers ), F3 and B3 ( outer primers ), MgSO4 and DNA template. The reaction mixture during the initial denaturation at 94 ° C for 5 minutes in a heating block was placed, then the Bst DNA polymerase (4 units) was added and the reaction mixture was incubated for 90 minutes at 65 ° C. Then LAMP products were analyzed by 2% agarose gel electrophoresis and SYBR Green. Results: The results showed that amplification of DNA was carried out in positive control according to ladder-shaped smears in agarose gel electrophoresis and SYBR Green visual. Conclusion: Rapid amplification, simple operation, no need a thermocycler, low cost and easy detection is advantages of LAMP assay than other molecular methods for detection of SEB.

Staphylococcus aureus, Enterotoxins B, Detection, LAMP