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Focusing on the research and development and application of isothermal nucleic acid amplification technology
Establishment of a Fluorescence Detection Method for RAA, a Recombinase-Aided Isothermal Amplification Technique for Porcine Eperythrozoon—China Animal Health and Veterinary Medicine
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To establish an accurate and rapid detection method for *Eperythrozoon suis*, specific primers and probes were designed based on the conserved sequence of the α-enolase gene of *Eperythrozoon suis*. After screening and optimizing reaction conditions, a recombinase-mediated isothermal amplification (RAA) fluorescence detection method was developed. The results showed that the established method can specifically detect *Eperythrozoon suis* within 20 minutes at a constant temperature of 39°C. It exhibits no cross-reactivity with *Mycoplasma hyopneumoniae*, *Streptococcus suis*, *Escherichia coli*, classical swine fever virus, porcine circovirus type 2, African swine fever virus, or healthy pig blood. The method’s limit of detection is as low as 1 copy/μL. When applied to 15 clinical samples, the method yielded a positive rate of 60%, which was consistent with the results obtained by real-time fluorescent quantitative PCR. The fluorescence RAA detection method for *Eperythrozoon suis* established in this study demonstrates good sensitivity and specificity, and its simple operation makes it suitable for rapid clinical detection of *Eperythrozoon suis* infection in pigs.
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