Development of Triplex RT-RAA Rapid Detection Technique for Important Swine Coronaviruses
Zheng Haotian
Zhang Biao
Cong Jiahao
Ling Xianghui
Yuan Wenju
Huang Huihui
Liu Guanshang
Chen Zhaojiang
Shi Hanxue
Wu Yuxuan
Guo Yitong
Yuan Jin
Wan Bo
Zheng Guanmin
Zhang Gaiping
Zhang Yuhang
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.
Keywords：Porcine epidemic diarrhea virusTransmissible gastroenteritis virusPorcine deltacoronavirusRecombinase-aided amplificationMultiplex detection
Publication Date：2026-08-15
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：11( 125-135 )