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Background   Staphylococcus aureus  is an important human pathogen causing a variety of life-threatening diseases. Rapid and accurate detection of  Staphylococcus aureus  is a necessity for prevention of outbreaks caused by this pathogen. PCR is a useful tool for rapid detection of foodborne pathogens, however, its inability to differentiate DNA from dead cells and live cells in amplification severely limits its application in pathogen detection. The aim of this study was to develop an improved assay was developed by incorporating the sample treatments with a surfactant and propidium monoazide (PMA) in qPCR for detection of viable  S. aureus  cells.    Results  The cell toxic effect testing with the two surfactants showed that the viability of  S. aureus  was virtually not affected by the treatment with 0.5% triton x-100 or 0.025% sarkosyl. Triton x-100 was coupled with PMA for sample treatments for detection of viable  S. aureus  cells in artificially contaminated milk. The qPCR results indicated that the assay reached high an amplification efficiency of 98.44% and the live  S. aureus  cells were accurately detected from the triton-treated spiked milk samples by the PMA-qPCR assay.    Conclusions  The qPCR assay combined with treatments of PMA and surfactants offers a sensitive and accurate means for detection of viable  S. aureus  cells. Cell toxic effect testing with the two surfactants showed that the viability of  S. aureus  was virtually not affected by the treatment with 0.5% triton x-100 or 0.025% sarkosyl. The information on sample treatment with surfactants to improve the dead cell DNA removal efficiency in qPCR by increasing PMA’s permeability to dead cells can be used for other pathogens, especially for Gram-positive bacteria.

An improved assay for rapid detection of viable Staphylococcus aureus cells by incorporating surfactant and PMA treatments in qPCR