Molecular Characterization and SYBR Green Ⅰ-Based Quantitative PCR for Duck Hepatitis Virus Type 1

Molecular Characterization and SYBR Green Ⅰ-Based Quantitative PCR for Duck Hepatitis Virus

Type 1

LUO Yu-jun;ZHANG Gui-hong;XU Xiao-qin;CHEN Jian-hong;LIAO Ming

【期刊名称】《农业科学学报（英文版）》 【年(卷),期】2008(007)009

【摘要】To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain,real-time quantitative polyrnerase chain reaction (RTQ-PCR) assay based on SYBR Green Ⅰ technology was developed to target 3D gene of DHV-1.Comparative sequence analysis showed that the genome has a typical picornarivus genetic organization,and strain DHV-1 R genetic organaiztion is 5' untranslated region (UTR)-VPO-VP3-VP1-2A1-2A2-2B-2C-3A-3B-3C-3D-3'

UTR,DHV-1

R

has

close

relationship with Parechovirus,and has 95.1-99.1% nucleotide sequence identity with other DHV-1 strains.Based on the DHV-1 sequences in GenBank,three pairs of specific primers were designed to amplify DHV-1 using real-time PCR.The results showed that real-time PCR Tm value is 85.6℃ and the real-time PCR provides a broad dynamic range,detecting from 102 to 109 copies of DHV-1 cDNA per reaction.No cross-reactions were found in specimens containing DPV,AIV and NDV.It is concluded that DHV-1 belongs to a new group of the family Picornaviridae that may form a separate genus most closely related to the genus

Parechovirus.All results showed that the real-time PCR has high sensitivity and specificity to detect DHV-1 using SYBR Green Ⅰ dissociation curve analysis,isolates can be distinguished by their melting temperature.These methods are rapid,sensitive,and reliable,and can be readily adapted for detection of DHV-1 from other clinical samples. 【总页数】7页(1140-1146)

【关键词】duck hepatitis virus type 1,complete genome,real-time RT-PCR

【作者】LUO Yu-jun;ZHANG Gui-hong;XU Xiao-qin;CHEN Jian-hong;LIAO Ming

【作者单位】College of Veterinary Medicine,South China Agricultural University,Guangzhou Science,Foshan

510642,P.R.China;College

of

Life

and of

University,Guangzhou 528231,P.R.China;College

Veterinary Medicine,South China Agricultural University,Guangzhou 510642,P.R.China;College

of

Veterinary

Medicine,South

China

Agricultural University,Guangzhou 510642,P.R.China;College of Life and Science,Foshan

University,Guangzhou

528231,P.R.China;College

of

Veterinary Medicine,South China Agricultural University,Guangzhou 510642,P.R.China 【正文语种】中文 【中图分类】S8 【文献来源】

https://www.zhangqiaokeyan.com/academic-journal-cn_journal-integrative-agriculture_thesis/020126273621.html 【相关文献】

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