Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
重组酶介导等温核酸扩增方法快速检测土壤沙门氏菌_郭雨
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Salmonella is a pathogenic bacterium capable of causing diseases in both humans and animals, and it ranks among the most common foodborne bacteria. Salmonella present in soil can contaminate plants like vegetables, subsequently entering the human body and triggering foodborne illnesses. However, due to the complex nature of soil and interference from other microorganisms, rapidly identifying whether soil has been contaminated with Salmonella remains a significant challenge. **Objective:** To develop a rapid and highly sensitive real-time recombinase-aided amplification (RT-RAA) method for detecting Salmonella in soil samples. **Methods:** Specific primers and probes targeting the invA gene sequence of Salmonella were designed to construct a recombinant plasmid containing the invA gene fragment for testing. The sensitivity of the RT-RAA method was evaluated using this plasmid. Additionally, the specificity of the method was assessed by employing genomic DNA templates from Salmonella, Escherichia coli, Shigella flexneri, and Staphylococcus aureus. Finally, the RT-RAA method was applied to detect Salmonella in soil samples collected from tomato and ginger plants, with results further validated using conventional plate culture methods. **Results:** The RT-RAA method successfully detected the invA gene fragment in the recombinant plasmid within 20 minutes at 39°C, achieving a detection limit as low as 10 plasmid copies per reaction. Importantly, no cross-reactivity was observed with Escherichia coli, Shigella flexneri, or Staphylococcus aureus. Soil sample analysis revealed that the tomato soil was indeed contaminated with Salmonella, while the ginger soil remained free of the pathogen—results that perfectly aligned with those obtained via plate culture. **Conclusion:** The RT-RAA method demonstrates high sensitivity and strong specificity, making it an effective tool for the rapid detection of Salmonella contamination in soil samples.
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