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Development of One-Step Reverse Transcriptase–Polymerase Chain Reaction to Detect Duck Hepatitis Virus Type 1
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Min-Chul | - |
dc.contributor.author | Kwon, Yong-Kuk | - |
dc.contributor.author | Joh, Seong-Joon | - |
dc.contributor.author | Kwon, Jun-Hun | - |
dc.contributor.author | Kim, Jae-Hong | - |
dc.contributor.author | Kim, Sun-Joong | - |
dc.date.accessioned | 2009-08-11T23:09:13Z | - |
dc.date.available | 2009-08-11T23:09:13Z | - |
dc.date.issued | 2007 | - |
dc.identifier.citation | Avian Dis. 51:540-545. 2007 | en |
dc.identifier.issn | 0005-2086 | - |
dc.identifier.uri | https://hdl.handle.net/10371/6982 | - |
dc.description.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. | en |
dc.description.sponsorship | We thank Hyuk-Man Kwon for propagation of DHV-1 samples and other viruses. This work was supported financially by the National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, Republic of Korea. | en |
dc.language.iso | en | - |
dc.publisher | American Association of Avian Pathologists | en |
dc.subject | duck hepatitis virus (DHV) type 1 (DHV-1) | en |
dc.subject | one-step RT-PCR | en |
dc.subject | RNA-dependent RNA polymerase | en |
dc.title | Development of One-Step Reverse Transcriptase–Polymerase Chain Reaction to Detect Duck Hepatitis Virus Type 1 | en |
dc.type | Article | en |
dc.contributor.AlternativeAuthor | 김민철 | - |
dc.contributor.AlternativeAuthor | 권용국 | - |
dc.contributor.AlternativeAuthor | 조성준 | - |
dc.contributor.AlternativeAuthor | 권준헌 | - |
dc.contributor.AlternativeAuthor | 김재홍 | - |
dc.contributor.AlternativeAuthor | 김선중 | - |
dc.identifier.doi | 10.1637/0005-2086(2007)51[540:DOORTC]2.0.CO;2 | - |
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