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Development of One-Step Reverse Transcriptase–Polymerase Chain Reaction to Detect Duck Hepatitis Virus Type 1

Cited 27 time in Web of Science Cited 0 time in Scopus
Authors
Kim, Min-Chul; Kwon, Yong-Kuk; Joh, Seong-Joon; Kwon, Jun-Hun; Kim, Jae-Hong; Kim, Sun-Joong
Issue Date
2007
Publisher
American Association of Avian Pathologists
Citation
Avian Dis. 51:540-545. 2007
Keywords
duck hepatitis virus (DHV) type 1 (DHV-1)one-step RT-PCRRNA-dependent RNA polymerase
Abstract
A one-step reverse transcriptase–polymerase chain reaction (RT-PCR) method was developed and optimized for the detection of duck hepatitis virus type 1 (DHV-1) using the Viral Gene-spin™ viral DNA/RNA extraction kit. A pair of DHV-1–specific primers was designed against the gene encoding RNA-dependent RNA polymerase (3D gene). Using RNA prepared from duckling liver samples infected with two reference and seven Korean field isolates of DHV-1, one-step RT-PCR with DHV-1–specific primers amplified a 467-bp fragment. Under the same conditions, no amplification was observed for 14 other avian pathogenic viruses and bacteria. Using RNA prepared from serial dilutions of the DHV-1 with the supernatant of the uninfected duckling liver homogenate (10% w/v), the one-step RT-PCR assay was found to be sensitive to 10 50% egg lethal dose (ELD50) 0.1 ml−1 of DHV-1. Furthermore, this method detected DHV-1 from the livers and allantoic fluid of duck embryos dying before 3 days postinoculation (PI) and of chicken embryos that were chilled at 3 days PI. Therefore, this one-step RT-PCR method is rapid, sensitive, and reliable, and can be readily adapted for detection of DHV-1 from other clinical samples.
ISSN
0005-2086
Language
English
URI
https://hdl.handle.net/10371/6982
DOI
https://doi.org/10.1637/0005-2086(2007)51[540:DOORTC]2.0.CO;2
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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