毛蚶中肠道病毒检测方法的建立

Development of Method for the Detection of Enteroviruses from Clams

本文建立了检测毛蚶中肠道病毒的方法。病毒在pH4.5时吸附于毛蚶组织,离心后用pH9.5甘氨酸盐水将病毒从毛蚶组织中洗脱出来,并用0.0005mol/L三氯化铝代替Johnson法中的Cat—Floc,达到去除细胞毒性物质的目的。经过再一次离心,利用牛肉浸膏在pH3.5—4.0时凝聚并吸附病毒的特性使洗脱液中的病毒得到浓缩。离心后沉淀溶解于0.1mol/L磷酸氢二钠溶液。用本法提取毛蚶中脊髓灰质炎病毒,当病毒含量为每克组织3.5×10~5至35PFU时,平均回收率为71.2％。

A procedure for concentrating and recovering enteroviruses from clams (Arca( Anadara)subcrenata Lischke)has been developed and evaluated. Viruses were efficiently adsorbed to homogenized clam tissue by adjusting the homogenate to pH4.5. After low-speed centrifugation, the viruses were eluted from the sediment with pH 9.5 0.05mol/L glycine-saline. Instead of using Cat-Floc in Johnson's method, AlCl_3 was used to remove cytotoxity from clam tissue homogenates. After low-speed centrifugation, viruses were concentrated by mixing the glycine supernatant with beef extract solution and by adjusting pH to 3.5—4.0. After another centrifugation, the pellet was resuspended in a small volume of phosphate buffer. This technique successfully recovered an average of 71.2％ of the poliovirus added to clam homogenates at levels of 35 to 3.5×10~5 PFU per 1 g.