关键词: 猪流行性腹泻病毒, 猪传染性胃肠炎病毒, 猪δ冠状病毒, 重组酶介导等温扩增, 多重检测

Abstract: To address the challenge of differentiating the major pathogens of viral diarrhea in piglets that cause similar clinical symptoms，including porcine epidemic diarrhea virus（PEDV），transmissible gastroenteritis virus（TGEV），and porcine deltacoronavirus（PDCoV），the highly conserved RdRp genes of these three viruses were used as the target. Specific primers suitable for recombinase‐aided amplification（RAA） were designed and screened，and a triplex RAA assay for the simultaneous detection of PEDV，TGEV，and PDCoV was established.The results showed that after reaction at 37 ℃ for 10 min，specific amplification bands of 137，289，170 bp could be detected by agarose gel electrophoresis.The detection limits for PEDV，TGEV，and PDCoV were 8.9，1.1，7.5 copies/μL，respectively，and no cross‐reactivity was observed with porcine reproductive and respiratory syndrome virus（PRRSV），porcine astrovirus（PAstV），or classical swine fever virus（CSFV）.When applied to 93 clinical samples for parallel comparative testing，the overall detection rate was 5.38%.Compared with the results of previously reported singleplex quantitative PCR assays for PEDV，TGEV，and PDCoV，the coincidence rate reached 100%，demonstrating good clinical application reliability. In summary，this method offers an accurate and efficient new detection tool for the rapid field differentiation of porcine enteric coronaviruses.

Key words: Porcine epidemic diarrhea virus, Transmissible gastroenteritis virus, Porcine deltacoronavirus, Recombinase‐aided amplification, Multiplex detection