Sobsey M D, Carrick R J, Jensen H R
Appl Environ Microbiol. 1978 Jul;36(1):121-8. doi: 10.1128/aem.36.1.121-128.1978.
New and improved methods for concentrating enteroviruses, reoviruses, and adenoviruses from oysters have been developed and evaluated. Viruses are efficiently adsorbed to homogenized oyster meat by adjusting the homogenate to pH 5.0 and a conductivity of less than or equal to 2,000 mg of NaCl per liter. After low-speed centrifugation, the virus-free supernatant is discarded and the viruses are eluted from the sedimented oyster solids with pH 7.5 glycine-NaCl having a conductivity of 8,000 mg of NaCl per liter. The oyster solids are removed by low-speed centrifugation and filtration, and the viruses in the filtered supernatant are concentrated to a small volume by either ultrafiltration or acid precipitation at pH 4.5. The concentrate is treated with antibiotics and inoculated into cell cultures for virus isolation and quantitation. When these methods were tested with oysters experimentally contaminated with polioviruses, reoviruses, and adenoviruses, recovery efficiencies averaged about 46%. With the exception of virus assay and quantitation, these methods are simple and inexpensive enough to be done in typical shellfish microbiology laboratories.
已经开发并评估了从牡蛎中浓缩肠道病毒、呼肠孤病毒和腺病毒的新的改进方法。通过将匀浆调节至pH 5.0且电导率小于或等于每升2000毫克氯化钠,病毒能有效地吸附到匀浆化的牡蛎肉上。低速离心后,弃去无病毒的上清液,并用pH 7.5、电导率为每升8000毫克氯化钠的甘氨酸 - 氯化钠溶液从沉淀的牡蛎固体中洗脱病毒。通过低速离心和过滤去除牡蛎固体,过滤后的上清液中的病毒通过超滤或在pH 4.5下酸沉淀浓缩至小体积。浓缩物用抗生素处理,然后接种到细胞培养物中进行病毒分离和定量。当用实验感染脊髓灰质炎病毒、呼肠孤病毒和腺病毒的牡蛎测试这些方法时,回收效率平均约为46%。除病毒检测和定量外,这些方法简单且成本低廉,足以在典型的贝类微生物实验室中进行。