肝胰腺细小病毒和斑节对虾杆状病毒的PCR复合检测  

The Detection of Hepatopancreatic Parvovirus and Monodon Baculovirus(MBV)by Multiplex Polymerase Chain Reaction

作　　者：刘鸿玲[1] 刘敏[1] 闫冬春[1] 

机构地区：[1]鲁东大学生命科学学院,山东烟台264025

出　　处：《水产科学》2011年第8期485-490,共6页Fisheries Science

基　　金：山东省高等学校科技计划项目(J09LC01);山东省高等学校优秀骨干教师国际合作培养项目(2007)

摘　　要：肝胰腺细小病毒和斑节对虾杆状病毒是常见的2种DNA对虾病毒,在虾类养殖业中经常会出现混合感染。因此,建立这2种病毒的复合检测方法显得尤为重要。根据基因库中肝胰腺细小病毒和斑节对虾杆状病毒的保守序列,设计了肝胰腺细小病毒和斑节对虾杆状病毒的特异性引物,对这2种病毒PCR复合检测的反应条件和试剂浓度进行优化,退火温度55℃,主要试剂Mg2+终浓度为3mmol/L,dNTP终浓度为0.4 mmol/L,引物终浓度为0.8μmol/L。进一步比较了在优化后的PCR反应体系下,检测单种病毒和同时检测这2种病毒的灵敏度差异。A method to detect Hepatopancreatic parvovirus（HPV） and Monodon baculovirus（MBV）as two conventional DNA viruses in penaeid,often combined infection,is needed.Two pairs of specific primers were designed based on the conserved sequence of HPV and MBV in the GeneBank,and the reaction conditions and reagent concentration were optimized for PCR detection of HPV and MBV.The optimal detection of the two viruses was observed at the following conditions：annealing temperature of 55 ℃,the final Mg2＋concentration of 3 mmol/L,the final dNTP concentration of 0.4 mmol/L and the final primer concentration of 0.8 μmol/L.Moreover,the differences in sensitivity for one-virus-testing and two-virus-testing were compared under the optimal condition.

关 键 词：肝胰腺细小病毒 斑节对虾杆状病毒 PCR复合检测 

分 类 号：S917[农业科学—水产科学]