A simple and rapid molecular detection of Staphylococcus aureus strain B using multiplex PCR

Abstract

Background: Staphylococcucs aureus (S. aureus) is one of the most important and pathogenic species from
Staphylococcaceae family. S. aureus is capable produced several enterotoxins (SEs) that classified in different
groups from A-R.

Objective: The present study aimed to set up simple and rapid detection of S. aureus strain B to develop a multiplex. polymerase chain reaction (PCR) assay for identification of genus and strain B of this bacteria.

Methods: In this work, after designing of specific primers for 23S rRNA and SEB, one step multiplex PCR was
carried out and the PCR product analyzed by agarose gel electrophoresis. The specificity of this method was
determined using different strains of S. aureus A (SEA), S.aureus C (SEC), Bacillus subtilis, Bacillus cereus, and
Listeria monocytogenes. Also, the sensitivity was distinguished by serial dilution of DNA genome from S. aureus.

Results: The PCR product was indicated at 226 and 1,250 bp specific amplified fragments according to amplification of SEB and 23SrRNA, respectively. The sensitivity of this method determined about 2 10³ CFU/ml and also the results showed that according to specific primers, the PCR is very specific.

Conclusion: This multiplex PCR assay can be used like a simple diagnostic test in clinical laboratories for identification of S. aureus.