Key result
The novel RT-PSR assay detected PEDV in 58.46% of clinical samples compared to 53.84% with conventional RT-PCR, demonstrating 10 times higher sensitivity than conventional RT-PCR.
Why the study?
Point-of-care testing in the field for PEDV is lacking due to the need for a simple, robust field-applicable test not requiring professional laboratory equipment.
The novel RT-PSR assay provides a rapid, highly sensitive, and specific visual detection method for porcine epidemic diarrhea virus, suitable for field use.
BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is a major etiological agent of porcine epidemic diarrhea around the world. Point-of-care testing in the field is lacking owing to the requirement for a simple, robust field applicable test that does not require professional laboratory equipment. The aim of this study was to establish a novel reverse transcription polymerase spiral reaction (RT-PSR) assay for the rapid detection of porcine epidemic diarrhea virus (PEDV). For the assay, a specific RT-PSR primer pair was designed against a conserved region in PEDV ORF3. RESULTS: The RT-PSR was optimized, and PEDV could be detected after a 50 min incubation at 62 °C, in addition to the 15 min required for reverse transcription. No cross-reaction with other porcine infectious viruses was observed. This new method for PEDV detection was 10 times more sensitive than the conventional reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates for 65 clinical samples using the new RT-PSR assay and the conventional RT-PCR assay were 58.46% (38/65) and 53.84% (35/65), respectively. In the RT-PSR assay, the addition of a mixture of dyes allowed a positive reaction to be directly observed by the naked eye. CONCLUSIONS: These results indicate that this RT-PSR assay is capable of accurately detecting PEDV, and has the advantages of high specificity and sensitivity for the detection of PEDV.
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Wang et al. (2019) studied Porcine epidemic diarrhea virus (PEDV) (n=65). Reverse transcription polymerase spiral reaction (RT-PSR) vs. Conventional RT-PCR was evaluated on Positive detection rate. The novel RT-PSR assay detected PEDV in 58.46% of clinical samples compared to 53.84% with conventional RT-PCR, demonstrating 10 times higher sensitivity than conventional RT-PCR.
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