摘要
在 27 ℃,pH 8.0,盐度 33条件下,用套式 PCR方法和斑点杂交方法对日本对虾( Penaeus japonicus)体内潜伏性感染 WSSV的方法进行了研究.对健康日本对虾注射不同稀释倍数的白斑综合症病毒粗提液,死亡率统计显示注射 10 4, 10 6倍稀释液组的日本对虾死亡率明显低于 10, 10 2, 10 3倍组,结合套式 PCR方法和斑点杂交方法进行病毒检测结果发现,仅 10, 10 2倍稀释液组斑点杂交和一步 PCR结果呈阳性; 10 3, 10 4, 10 6倍稀释液组经二步 PCR检测为阳性、斑点杂交呈阴性,该结果为建立或界定 WSSV在日本对虾体内的潜伏性感染提供了方法学上的依据.
Under conditions of 27 t, pH 8.0 and salinity 33, we studied the method of latent infection of white spot syndrome virus(WSSV) in Penaeus japonicus by nested-PCR and dot blot hybridization. Healthy Penaeus japonicus were injected with serially 10-fold diluted WSSV stored solution. Results show that accumulated mortality rates of shrimps which were injected with stored solution diluted 10 4, 10 6 times are more less than that of shrimps injected with solution diluted 10, 10 2, 10 3 times. After detection of nested-PCR and dot blot hybridization, only 10,10 2 groups show positive results by one-step PCR and dot blot hybridization. 10 3, 10 4, 10 6 groups are positive by two-step PCR , but they are negative by dot blot hybridization. This experiment provides methodological evidence for establishing or seeking boundaries of latent infection of WSSV in Penaeus japonicus .
出处
《海洋科学》
CAS
CSCD
北大核心
2003年第8期72-76,共5页
Marine Sciences
基金
国家973项目"环境胁迫对对虾抗病力的影响和人工调控"G1999012011号
国家杰出青年基金40025614号
中国科学院知识创新领域前沿项目200223106号
关键词
白斑综合症病毒
日本对虾
潜伏性感染
斑点杂交
套式PCR
Penaeus japonicus, Nested-PCR, Dot blot hybridization, White spot syndrome virus,Latent infection
