学术
SYBR Green Ⅰ实时定量PCR快速检测鸭瘟病毒的研究
汤承[1] 索化夷[1,3] 岳华[1] 杨发龙[1] 汤明[2]
[1]西南民族大学生命科学与技术学院,四川成都610041 [2]重庆动物卫生监督站,四川成都610041 [3]西南大学食品科学学院,四川成都610041
摘 要:
根据GenBank中鸭瘟病毒（DPV）UL6和UL7基因的保守序列，设计了一对特异性引物，扩增位于UL6和UL7基因第891-991位长度为101bp片断。以DPV标准强毒株DNA为模板，建立了SYBR Green Ⅰ实时荧光定量PCR检测鸭瘟病毒的方法，用该方法检测鸭瘟标准强毒、疫苗毒及野毒株均为阳性，检测其他受试的非鸭瘟病毒DNA均为阴性，对鸭瘟强毒和疫苗毒人工感染鸭胚尿囊液的检出率均为100％，对6个临床送检样本的检出率为6／6，与病毒分离鉴定结果一致；最小检出量为1．57×10^4拷贝／ul。[著者文摘]
文章出处:
《中国动物检疫》-2006年23卷9期 -25-27页
栏目信息:
分 类 号:
文章编号:
1005-944X（2006）09-0025-03
Establishment of SYBR Green Ⅰ Real-Time Fluorescent Quantitative PCR for Detection of Duck Plague Virus
Tang Cheng, Suo Hua-yi,Yue Hual,Yang Fa-long,Tang Ming,Huang Guo-jun （1.College of Life Science and Technology, Southwest University for Nationality ,Chengdu, 610041;2.Chongqing Veterinary Health Supervision Station,401147;3.College of Food Science,Southwest University,chongqing, 400715）
Abstract:
To develop a SYBR Green Ⅰ real-time quantitative PCR for detection of duck plague virus （DPV）,specific primers were designed by using Beacon Designer software according to the UL6 and UL7 gene sequence available in GenBank. The amplicon locates between 891-991 with a size of 101bp. With DNA obtained from standard virulent strain of DPV, a real-time quantitative PCR （DPV- SYBR Green Ⅰ PCR） was established successfully. The resuits showed this assay was specific and sensitive for DPV with a detection limit of 1.57×10^4 copies . The allantoic fluid obtained from duck embryo experimentally infected with f37 and C-KCE were detected using the assay and were all positive. When using the assay to detect clinical samples, the same results （6/6 positive） was obtained compared with virus isolation. The assay alsoproven to be specific,and the detection limit was upto 1.57×10^4 copies.[著者文摘]
Key words:
duck plague virus; UL6 and UL7 gene; SYBR Green Ⅰ; real-time PCR
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