Data Sheet 1_Development and evaluation of a recombinase-aided amplification-lateral flow dipstick assay for rapid and sensitive detection of Mycoplasma synoviae.zip

To further evaluate the clinical performance of our RAA-LFD-MS assay, a total of 100 clinical specimens suspected of MS infection were tested in parallel using this newly established method, conventional polymerase chain reaction (PCR), and quantitative real-time PCR (qPCR).</p>Results<p>The RAA reaction was completed within 20 min at a constant temperature of 37 °C, and amplification products were visually read within 3–5 min using a lateral flow dipstick without sophisticated equipment.

The total detection time, including the DNA extraction step, is completed within 40 min. Methodological validation revealed that the RAA-LFD assay exhibited high specificity, with no cross-reactivity observed against other common avian pathogens. The assay also showed satisfactory repeatability and high sensitivity, with a limit of detection of 2.63×10⁰ copies/μL. In the clinical sample evaluation, the coincidence rate between RAA-LFD and PCR was 96%, while the agreement between RAA-LFD and qPCR reached 100%.</p>Conclusion<p>In summary, the established RAA-LFD assay is rapid, user-friendly, highly specific, and sensitive.

To further improve the RAA-LFD-MS assay for routine clinical and field use, future efforts should prioritize the optimization of rapid DNA extraction, accessible temperature-control devices, and reagent lyophilization.</p>