A novel, rapid, and sensitive real-time PCR assay for cost-effective detection and quantification of Staphylococcus aureus in food samples with the ZEN™ doublequenched probe chemistry
2019
Salihah, N. T. | Hossain, M. M. | Abdul Hamid, M. R. W. | Ahmed, M. U.
Staphylococcus aureus is a prevalent food pathogen. The pathogen is very dangerous as it could survive high cooking temperature and produce heat-stable toxins. Therefore, a fast, sensitive and secure culture-free alternative to detect and correctly quantify S. aureus in food samples to prevent food poisoning associated with it is needed. In the present work, the development of a comparatively inexpensive real-time qPCR assay based on the ZENTM probe chemistry by targeting the thermonuclease gene for a fast, sensitive, cost-effective, and direct quantitative detection of S. aureus in real food samples is reported. The novel assay showed high specificity and sensitivity with the limit of detection (LoD) of as low as 1 copy /reaction. The quantification range varies between 1×10¹ and 1×10 7 copies/reaction. This method was able to accurately quantify S. aureus from milk sample with a relative accuracy of 75.88 to 120%. The novel assay successfully detected the presence of as low as 50 copies/mL or copies/g of S. aureus in real food matrices.
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