Rapid and Simple Detection of Viable Foodborne Pathogen Staphylococcus aureus.

Caiyan Liu,Cuiping Ma,Mengyuan Wang,Zonghua Wang,Chao Shi,Mengzhe Li

doi:10.3389/fchem.2019.00124

Abstract

Staphylococcus aureus (S. aureus) contamination in food safety has become a worldwide health problem. In this work, we utilized RNA one-step detection of denaturation bubble-mediated Strand Exchange Amplification (SEA) method to realize the detection of viable foodborne pathogen S. aureus. A pair of S. aureus specific primers were designed for the SEA reaction by targeting hypervariable V2 region of 16S rDNA and the amplification reaction was finished about 1 h. The results of amplification reaction could be observed by the naked eyes with a significant color change from light yellow to red to realize the colorimetric detection of S. aureus. Therefore, there only required an isothermal water bath, which was very popular for areas with limited resources. In real sample testing, although the SEA detection was so time-saving compared with the traditional plating method, the SEA method showed great consistency with the traditional plating method. In view of the above-described advantages, we provided a simple, rapid and equipment-free detection method, which had a great potential on ponit-of-care testing (POCT) application. Our method reported here will also provide a POCT detection platform for other food-borne pathogens in food, even pathogenic bacteria from other fields.

Highlights

Staphylococcus aureus (S. aureus), a facultative anaerobic Gram-positive coccus, can greatly threaten our health due to a combination of toxin-mediated virulence, invasiveness, and antibiotic resistance (Oliveira et al, 2011)
The results showed that the detection limit of the Strand Exchange Amplification (SEA) method on S. aureus was 100 cfu/g (Figure S3), which met the requirements that S. aureus must not be detected out in foods
A rapid and simple SEA method for the detection of S. aureus was developed in this work

Summary

Introduction

Staphylococcus aureus (S. aureus), a facultative anaerobic Gram-positive coccus, can greatly threaten our health due to a combination of toxin-mediated virulence, invasiveness, and antibiotic resistance (Oliveira et al, 2011). S. aureus is the main cause of nosocomial infections and community-acquired diseases, including deep-seated, endocarditis, abscesses and bacteria, which lead to toxic and septic shock syndromes (Abdalhai et al, 2014). S. aureus widely exists in the air, water, dust, human and animal excretions, which makes the food much easier to become contaminated (Yu et al, 2016). It has been reported that S. aureus contamination is a worldwide health problem. In the United States, nearly half a million hospitalizations and 50,000 deaths occur resulting from S. aureus each year (Schlecht et al, 2015). 11 outbreaks of Staphylococcal food poisoning were reported between 2006 and 2009 in Shenzhen, China, which ranked the second

Methods

Results

Conclusion