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洗脱标记物在脊髓灰质炎病毒株型内鉴定中的应用。

Use of elution marker for the intratypic characterization of poliovirus strains.

作者信息

Szöllõsy E, Mucsi I, Pálmai M

出版信息

Acta Microbiol Acad Sci Hung. 1976;23(4):301-8.

PMID:193364
Abstract

Elution marker was used for intratypic characterization of poliovirus strains with Al(OH)3 gel as adsorbent. The virion suspensions to be tested were partially purified by chromatography and labelled with 32P. In the labelled preparations of wild virus strains practically all radioactivity was found in virus-specific bond, whereas in those of the vaccine strains and isolates of vaccine origin a considerable, but variable, proportion of the activity was bound to residual cell components. For this reason, the EC50 value (i.e. the phosphate molarity corresponding to the 50% adsorption equilibrium of virions), for the vaccine and vaccine-like strains showed a wide scattering. The activity bound to cell components was removable from the gel with 0.005 M phosphate buffer, whereas the elution maxima for all the poliovirus strains examined so far were over 0.02 M. The elution marker was calculated for 25 type-2 and 30 type-3 strains, all isolated during or soon after vaccination periods from cases suspect of poliomyelitis, both with and without taking into account the cell-bound activity. In the latter case, the EC50 values agreed with that for the corresponding reference vaccine strain much better than in the former; furthermore, the type-2 reference vaccine strain and the type-2 isolates, indistinguishable from the wild reference strain by the original method, could easily be differentiated on the basis of the corrected EC50 value. The possibility that the lack of an appreciable proportion of cell-bound radioactivity may reflect the pathogenicity of poliovirus strains is discussed.

摘要

以氢氧化铝凝胶为吸附剂,使用洗脱标记物对脊髓灰质炎病毒株进行型内鉴定。待检测的病毒粒子悬液通过色谱法进行部分纯化,并用32P进行标记。在野生病毒株的标记制剂中,几乎所有放射性都存在于病毒特异性结合物中,而在疫苗株和疫苗源分离株的制剂中,相当一部分但可变比例的活性与残留细胞成分结合。因此,疫苗株和疫苗样株的EC50值(即对应病毒粒子50%吸附平衡的磷酸盐摩尔浓度)显示出广泛的离散性。与细胞成分结合的活性可用0.005M磷酸盐缓冲液从凝胶中去除,而迄今为止检测的所有脊髓灰质炎病毒株的洗脱最大值均超过0.02M。对25株2型和30株3型病毒株计算洗脱标记物,这些病毒株均在疫苗接种期间或接种后不久从疑似脊髓灰质炎病例中分离得到,计算时既考虑了与细胞结合的活性,也未考虑该活性。在后一种情况下,EC50值与相应参考疫苗株的EC50值比在前一种情况下更吻合;此外,2型参考疫苗株和2型分离株,用原方法与野生参考株无法区分,根据校正后的EC50值可轻松区分。文中讨论了缺乏相当比例与细胞结合的放射性可能反映脊髓灰质炎病毒株致病性的可能性。

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