Key result
The RT-RAA assay detected PEDV in clinical tissue samples with a 24.19% positive rate, matching the performance of RT-qPCR, and demonstrated high specificity without cross-reactivity.
Why the study?
PEDV causes high mortality and economic losses in swine, but there was no report on PEDV detection using RT-RAA to overcome limitations of existing methods.
Does RT-RAA accurately and rapidly detect PEDV in clinical tissue samples compared to RT-qPCR?
Does RT-RAA accurately and rapidly detect PEDV in clinical tissue samples compared to RT-qPCR?
RT-RAA provides a rapid, sensitive, and highly specific method for the onsite molecular diagnosis of PEDV in swine.
This is the authors' abstract. We don't add key points for this paper.
Porcine epidemic diarrhea virus (PEDV) is a coronavirus currently widespread worldwide in the swine industry. Since PEDV was discovered in China in 1984, it has caused huge economic losses in the swine industry. PEDV can infect pigs of all ages, but piglets have the highest infection with a death rate as high as 100%, and the clinical symptoms are watery diarrhea, vomiting, and dehydration. At present, there is not any report on PEDV detection by RT-RAA. In this study, we developed an isothermal amplification technology by using reverse-transcription recombinase-aided amplification assay (RT-RAA) combined with portable instruments to achieve a molecular diagnosis of PEDV in clinical samples from China. By designing a pair of RT-RAA primers and probes based on the PEDV N gene, this method breaks the limitations of existing detection methods. The assay time was within 30 min at 41 °C and can detect as few as 10 copies of PEDV DNA molecules per reaction. Sixty-two clinical tissue samples were detected by RT-qPCR and RT-RAA. The positive and negative rates for the two methods were 24.19% and 75.81%, respectively. Specificity assay showed that the RT-RAA had specifically detected PEDV and was not reactive for porcine parvovirus (PPV), transmissible gastroenteritis virus (TGEV), porcine circovirus type 2 (PCV2), porcine pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), swine flu virus (SIV), or porcine Japanese encephalitis virus (JEV). The results suggested that RT-RAA had a strong specificity and high detection sensitivity when combined with a portable instrument to complete the detection under a constant temperature of 30 min, which are more suitable for preventing and controlling PEDV onsite in China.
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RT-RAA assay enables rapid PEDV detection in swine; leaves open prospective validation before routine diagnostic use.
Wu et al. (2022) studied Porcine Epidemic Diarrhea Virus (n=62). RT-RAA assay vs. RT-qPCR was evaluated on PEDV detection rate. The RT-RAA assay detected PEDV in clinical tissue samples with a 24.19% positive rate, matching the performance of RT-qPCR, and demonstrated high specificity without cross-reactivity.
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