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Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus.
Cao, Wei W; He, Dong S; Chen, Zhen J; Zuo, Yu Z; Chen, Xun; Chang, Yan L; Zhang, Zhi G; Ye, Lei; Shi, Lei.
  • Cao WW; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • He DS; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Chen ZJ; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Zuo YZ; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Chen X; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Chang YL; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Zhang ZG; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Ye L; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
  • Shi L; Institute of Food Safety and Nutrition, Jinan University, Guangzhou, China (Cao, Z.J. Chen, X. Chen, Chang, Shi, Ye); College of Veterinary Medicine, South China Agricultural University, Guangzhou, China (He); College of Animal Science and Technology, Agricultural University of Hebei, Baoding, China
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.
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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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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