Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus.
J Vet Diagn Invest
; 32(4): 572-576, 2020 Jul.
Article
in English
| MEDLINE | ID: covidwho-823306
ABSTRACT
Porcine epidemic diarrhea, a disease caused by porcine epidemic diarrhea virus (PEDV), results in large economic losses to the global swine industry. To manage this disease effectively, it is essential to detect PEDV early and accurately. We developed a sensitive and accurate droplet digital PCR (ddPCR) assay to detect PEDV. The optimal primer-to-probe concentration and melting temperature were identified as 300200 nM and 59.2°C, respectively. The specificity of the ddPCR assay was confirmed by negative test results for common swine pathogens. The detection limit for the ddPCR was 0.26 copies/µL, which is a 5.7-fold increase in sensitivity compared to that of real-time PCR (rtPCR). Both ddPCR and rtPCR assays exhibited good linearity, although ddPCR provided higher sensitivity for clinical detection compared to that of rtPCR. Our ddPCR methodology provides a promising tool for evaluating the PEDV viral load when used for clinical testing, particularly for detecting samples with low-copy viral loads.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Swine Diseases
/
Polymerase Chain Reaction
/
Coronavirus Infections
/
Viral Load
/
Porcine epidemic diarrhea virus
Type of study:
Diagnostic study
/
Experimental Studies
/
Prognostic study
Limits:
Animals
Language:
English
Journal:
J Vet Diagn Invest
Journal subject:
Veterinary Medicine
Year:
2020
Document Type:
Article
Affiliation country:
1040638720924753
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