Development of one-step reverse transcriptase-polymerase chain reaction to detect duck hepatitis virus type 1. Export

@article{citeulike:1676059, 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 DHV1-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.}, address = {National Veterinary Research and Quarantine Service, 480 Anyang 6 dong, Manan-gu, Anyang, Gyeonggi-do, 420-824, Republic of Korea.}, author = {Kim, M. C. and Kwon, Y. K. and Joh, S. J. and Kwon, J. H. and Kim, J. H. and Kim, S. J.}, citeulike-article-id = {1676059}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/17626480}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=17626480}, issn = {0005-2086}, journal = {Avian Dis}, keywords = {detection, duck-hepatitis-virus}, month = {June}, number = {2}, pages = {540--545}, posted-at = {2007-09-19 14:50:04}, priority = {2}, title = {Development of one-step reverse transcriptase-polymerase chain reaction to detect duck hepatitis virus type 1.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/17626480}, volume = {51}, year = {2007} }

TY - JOUR ID - citeulike:1676059 L3 - citeulike-article-id:1676059 N2 - 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 DHV1-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. IS - 2 JF - Avian Dis SN - 0005-2086 AD - National Veterinary Research and Quarantine Service, 480 Anyang 6 dong, Manan-gu, Anyang, Gyeonggi-do, 420-824, Republic of Korea. EP - 545 TI - Development of one-step reverse transcriptase-polymerase chain reaction to detect duck hepatitis virus type 1. VL - 51 SP - 540 KW - detection KW - duck-hepatitis-virus AU - Kim, MC AU - Kwon, YK AU - Joh, SJ AU - Kwon, JH AU - Kim, JH AU - Kim, SJ PY - 2007/06// UR - http://view.ncbi.nlm.nih.gov/pubmed/17626480 ER -