Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
Rapid Detection of Salmonella in Soil by Isothermal Nucleic Acid Amplification Assay Mediated by Recombinase—Guo Yu
-
File size: 722.4KB
Salmonella is a pathogenic bacterium that can cause diseases in both humans and animals, and it is one of the most common foodborne bacteria. Salmonella present in soil can enter the human body through vegetables and other plants, leading to food poisoning. However, due to the complex nature of soil and interference from other microorganisms, rapidly identifying whether soil is contaminated with Salmonella remains a significant challenge. [Objective] To develop a rapid and sensitive real-time recombinase-aided amplification (RT-RAA) method for the detection of Salmonella in soil. [Methods] Specific primers and probes were designed targeting the invA gene sequence of Salmonella, and a recombinant plasmid containing the invA gene fragment to be tested was constructed. The sensitivity of the RT-RAA method was evaluated using this recombinant plasmid. The specificity of the RT-RAA method was assessed by using genomic DNA from Salmonella, Escherichia coli, Shigella flexneri, and Staphylococcus aureus as templates. The RT-RAA method was applied to detect Salmonella in soil samples from tomato and ginger fields, and the results were validated using conventional plate culture methods. [Results] The RT-RAA method can detect the invA gene fragment in recombinant plasmids, yielding results within 20 minutes at 39°C. The minimum detection limit was 10 plasmid copies per reaction, and no cross-reactivity was observed with Escherichia coli, Shigella flexneri, or Staphylococcus aureus. RT-RAA detection of soil DNA samples revealed that the tomato soil was contaminated with Salmonella, whereas the ginger soil was not, consistent with the results obtained by plate culture. [Conclusion] The RT-RAA method exhibits high sensitivity and strong specificity, making it suitable for the rapid detection of Salmonella contamination in soil.
Product Inquiry
Contact: qt@qt-bio.com
Product Consultation:+86-510-85385531
Technical support:+86-18921157475
Address: No. 97,Xingye Building B,Linghu Avenue,Xiwu District, Wuxi City
WeChat Official Account
Official Official Account
www.300.cn