Development and Application of a One-step Real-time TaqMan RT-PCR Assay for Detection of Duck Hepatitis Virus Type 1
作者单位：四川出入境检验检疫局 Avian Disease Research CenterCollege of Veterinary MedicineSichuan Agricultural University The Key Laboratory of Animal Disease and Human Health of Sichuan Province College of Life Science and Technology of Southwest University for Nationalities
会议名称：《第五届中国畜牧科技论坛》
会议日期：2011年
学科分类：090603[农学-临床兽医学] 09[农学] 0906[农学-兽医学]
基 金：supported by grants from the National Support Program of Science and Technology (2007Z06-017) New Century Excellent Talents program in University(NCET-04-0906/NCET- 06-0818) Scientific and Technological Innovation Major Project Funds in University(706050) Sichuan Province Basic Research Program(07JY029-016) Program for Key Disciplines Construction of Sichuan Province(SZD0418)
关 键 词：Duck hepatitis virus typel rRT-PCR Duck embryo Chicken embryo Distribution
摘 要：A one-step real-time RT-PCR assay(rRT-PCR) was developed for efficient detection of Duck hepatitis virus typel(DHV-1).A pair of specific primers was designed against the conserved region in the 3D gene that encodes the RNA dependent RNA polymerase with a single conserved TaqMan? *** detection limit of this assay was 10 viral genomic copies per reaction and it was highly specific to *** rRT-PCR assay was used to determine the distribution and concentration of DHV-1 virulent strain in duck embryos as well as the DHV-1 attenuated vaccine strain in chicken *** results revealed that the copy numbers of DHV-1 reached a peak in duck embryos and chicken embryos at 28-40h,44-56h post inoculation respectively. The comparative tests for ducklings infected artificially and clinical samples between neutralization test and rRT-PCR showed that the positive results of infected samples were the same,while the rRT-PCR method was more sensitive than neutralization test for detection of clinical *** rapid,sensitive and specific rRTPCR assay will be a powerful tool for detection of suspected cases of DHV-1,distribution pattern of DHV-1 in vivo and molecular epidemiological screening.