Johnson K M, Cooper R C, Straube D C
Appl Environ Microbiol. 1981 Apr;41(4):932-5. doi: 10.1128/aem.41.4.932-935.1981.
The most likely shellfish to be harvested if sportfishing is reinstated in San Francisco Bay is the Japanese cockle Tapes japonica. The virus levels present in these shellfish are unknown and need to be evaluated before the shellfish beds are open. Towards this end, a procedure for recovering and concentrating enteric viruses from these clams has been evaluated. Effective elution of poliovirus from clam tissues was found to occur with pH 9.5 glycine-buffered saline rather than with the pH 7.5 fluid utilized by other investigators on oysters. Poliovirus desorption was combined with Cat-Floc clarification to remove cytotoxicity from clam tissue homogenates. For assay purpose, viruses were concentrated by mixing the glycine supernatant with a beef extract solution, lowering the pH, and suspending the resulting floc in a small volume of phosphate buffer. This simple technique successfully recovered an average of 73% of the poliovirus added to clam homogenates at levels of 93 and 660 plaque-forming units per 100 g. Coxsackievirus B2 was isolated from clams exposed to raw sewage.
如果旧金山湾恢复休闲垂钓,最有可能被捕捞的贝类是日本鸟蛤。这些贝类中的病毒水平未知,在贝类养殖场开放之前需要进行评估。为此,已经评估了一种从这些蛤中回收和浓缩肠道病毒的方法。发现用pH 9.5甘氨酸缓冲盐水能有效地从蛤组织中洗脱脊髓灰质炎病毒,而不是其他研究人员用于牡蛎的pH 7.5溶液。脊髓灰质炎病毒解吸与Cat-Floc澄清相结合,以去除蛤组织匀浆中的细胞毒性。为了进行检测,通过将甘氨酸上清液与牛肉提取物溶液混合、降低pH值并将所得絮凝物悬浮在少量磷酸盐缓冲液中来浓缩病毒。这种简单的技术成功地回收了添加到蛤匀浆中的脊髓灰质炎病毒,平均回收率为73%,添加水平为每100克93和660个空斑形成单位。从暴露于未经处理污水的蛤中分离出了柯萨奇病毒B2。