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Use of an internal control in a quantitative RT-PCR assay for quantitation of porcine epidemic diarrhea virus shedding in pigs

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Authors
Song, Dae-Sub; Kang, Bo-Kyu; Lee, Sang-Sun; Yang, Jeong Sun; Moon, Hyoung-Joon; Oh, Jin-Sik; Ha, Gun-Woo; Jang, Yong-Suk; Park, BongKyun
Issue Date
2005-11-18
Publisher
Elsevier
Citation
J.Virol. Methods 133, 27-33
Keywords
Porcine epidemic diarrhea virusSheddingQuantitative RT-PCR
Abstract
Porcine epidemic diarrhea virus (PEDV), a member of the family Coronaviridae, has caused a devastating enteric disease in the Korean swine industry. Previously, the differences between virulent field PEDV strains and a Vero cell culture adapted PEDV DR13 strain were determined using restriction fragment length polymorphism analysis (RFLP), and PEDV shedding patterns in pigs were reported. In an extension to these studies, an internal control was constructed and quantitative analysis of virus shedding after oral inoculation was established. A parent field PEDV and a cell culture adapted PEDV DR13 were inoculated orally to colostrum-deprived 1-day-old piglets, commercial 2-week-old pigs, and sows (1–5 ml dose, 105.8–106.0 TCID50/0.1 ml). PEDV shedding was monitored every day and virus levels were measured using a quantitative reverse transcriptase polymerase chain reaction (RT-PCR) method. In fecal samples from experimentally-inoculated pigs, the level of virus excreted peaked at 2 days after oral inoculation and gradually decreased thereafter. In addition, PEDV from field specimens was quantified using the same RT-PCR assay to determine shedding viral load.

This suggests that measurement of PEDV shedding viral load in pigs, by quantitative RT-PCR, may be a useful tool for estimating the transmission potential of PEDV in the swine population.
ISSN
0166-0934
Language
English
URI
http://hdl.handle.net/10371/6903
DOI
https://doi.org/10.1016/j.jviromet.2005.10.021
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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