Ⅰ型鸭病毒性肝炎病毒和新型鸭肝炎病毒鉴别RT-PCR检测方法的建立  

Development of identify RT-PCR method for duck hepatitis virus type Ⅰ(DHV-Ⅰ) and new serotype duck hepatitis virus

作　　者：何冉娅[1] 罗玉均[1] 孙伟[2] 何逸民[1] 于淼[1] 张桂红[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]西南大学药学院,重庆400715

出　　处：《黑龙江畜牧兽医》2009年第3期14-16,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(30871877)

摘　　要：根据GenBank中公布的Ⅰ型鸭病毒性肝炎病毒和实验室分离到的Ⅰ型鸭病毒性肝炎ZJ株、Ⅰ型鸭病毒性肝炎R株所测序列（GenBank登录号分别为EU841005和EF585200）以及新型鸭肝炎病毒全基因组序列，应用PrimerPrimier5．0软件，在序列保守区域分别设计了2对鉴别引物，成功建立了Ⅰ型鸭病毒性肝炎病毒和新型鸭肝炎病毒的鉴别RT—PCR检测方法。结果表明：该方法能从Ⅰ型鸭病毒性肝炎病毒中扩增到与预期相符的471bp条带，从广东佛山分离到的新型鸭肝炎病毒FS株中扩增到705bp的条带，设计的2对引物能够特异性地检测出Ⅰ型鸭病毒性肝炎病毒和新型鸭肝炎病毒，特异性好；分别能够检测出模板含量为0．8ng／μL和1．0ng／μL的Ⅰ型鸭病毒性肝炎病毒和新型鸭肝炎病毒。因此，该方法可用于Ⅰ型鸭病毒性肝炎病毒和新型鸭病毒性肝炎临床样品的鉴别诊断和分子流行病学调查。In the study, identify RT - PCR method for duck hepatitis virus type Ⅰ（ DHV - Ⅰ ） and new serotype duck hepatitis virus was developed successfully using two pairs of primers designed with Primer Primier 5.0 software according to the complete genome sequence of DHV - Ⅰ and new serotype duck hepatitis virus deposited in GenBank （GenBank accession numer are EU841005 and EF585200 , respectively ）and ZJ strain and R strain in our lab. The result indicated that this method was shown to specifically amplify a 471 bp fragment from DHV -Ⅰ or a 705 bp fragment from new serotype duck hepatitis virus FS strain. Test on clinical samples showed that this method was highly specific and can detect DHV - Ⅰ to 0.8 ng/μL and new serotype duck hepatitis virus to 1 ng/μL in template respectively. Therefore, the identify RT - PCR could be used as an effective tool for diagnosis of DHV - Ⅰ and new serotype duck hepatitis virus in clinical samples, and molecule epidemiological investigations.

关 键 词：Ⅰ型鸭病毒性肝炎病毒 新型鸭肝炎病毒 鉴别RT—PCR 

分 类 号：S852.65[农业科学—基础兽医学]