Rapid Detection of Staphylococcus aureus in Food Using a Recombinase Polymerase Amplification-Based Assay
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Staphylococcus aureus (S. aureus) is of great importance and is a leading cause of food poisoning, which is a public health concern in terms of the frequency and seriousness of the disease. In the present study, RPA and real-time RPA assays were developed and validated to detect S. aureus with high sensitivity and specificity by targeting the nuc gene for the first time. The analytical sensitivity of real-time RPA was 102 copies/reaction, which was higher than the sensitivity of the real-time PCR method. The analysis time was reduced to 10 min, but this method was as reliable as real-time PCR. Furthermore, the potential use of RPA to detect S. aureus was validated with five different artificially contaminated foods. In conclusion, the RPA and real-time RPA assays developed here, similar to real-time PCR, are rapid and simple and exhibit with high sensitivity and specificity. These assays serve as efficient tools for the detection of S. aureus in less advanced laboratories and are suitable for point-of-care detection.
KeywordsStaphylococcus aureus RPA Real-time RPA Detection
This work was funded by the Natural Science Foundation of Hebei Province (C2017205129), the Research Project of General Administration of Quality Supervision, Inspection and Quarantine of China (2016IK107), Program for Young Top-notch Talents in Universities of Hebei Province (BJ2016033), Doctoral Foundation of Hebei Normal University (L2016B13), and One Hundred Person Project of Hebei Province (E2016100019).
Compliance with Ethical Standards
Conflict of Interest
Yunyun Geng declares that she has no conflict of interest. Siying Liu declares that she has no conflict of interest. Jinfeng Wang declares that she has no conflict of interest. Huizhu Nan declares that she has no conflict of interest. Libing Liu declares that he has no conflict of interest. Xiaoxia Sun declares that she has no conflict of interest. Danyu Li declares that she has no conflict of interest. Ming Liu declares that she has no conflict of interest. Jianchang Wang declares that he has no conflict of interest. Ke Tan declares that he has no conflict of interest.
This article does not contain any studies with human participants or animal performed by any of the authors.
- Brakstad OG, Aasbakk K, Maeland JA (1992) Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J Clin Microbiol 30:1654–1660Google Scholar
- Gao W, Huang H, Zhu P, Yan X, Fan J, Jiang J, Xu J (2018) Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish. Bioproc Biosyst Eng Publishied onlineGoogle Scholar
- International Organization for Standardization (2003) ISO 6888-3:2003. Microbiology of food and animal feeding stuffs–horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species): part 3: detection and MPN technique for low numbers. GenevaGoogle Scholar
- Klevens RM, Morrison MA, Nadle J, Petit S, Gershman K, Ray S, Harrison LH, Lynfield R, Dumyati G, Townes JM, Craig AS, Zell ER, Fosheim GE, McDougal LK, Carey RB, Fridkin SK (2007) Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 298:1763–1771CrossRefGoogle Scholar
- Murinda SE, Ibekwe AM, Zulkaffly S, Cruz A, Park S, Razak N, Paudzai FM, Ab Samad L, Baquir K, Muthaiyah K, Santiago B, Rusli A, Balkcom S (2014) Real-time isothermal detection of Shiga toxin-producing Escherichia coli using recombinase polymerase amplification. Foodborne Pathog Dis 11:529–536CrossRefGoogle Scholar
- National food safety standard of China (2016) Food microbiological examination: Staphylococcus aureus. GB4780.10-2016Google Scholar