建立白斑综合症病毒在日本对虾体内潜伏性感染的方法研究

在27℃，pH8．0，盐度33条件下，用套式PCR方法和王证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。

A Method Research of Establishing the Latent Infection of White Spot Syndrome Virus(WSSV) in Penaeus japonicus

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 .