Hovi T, Roivainen M
Department of Virology, National Public Health Institute, Helsinki, Finland.
J Clin Microbiol. 1989 Apr;27(4):709-15. doi: 10.1128/jcm.27.4.709-715.1989.
We have developed a new rapid test, the radiometric cytolysis inhibition assay (RACINA), for the determination of neutralizing poliovirus antibodies. HeLa cells prelabeled with 51Cr, [3H]leucine, or, preferentially, with [3H]uridine are used as sensitive quantitative indicators of residual infectious virus. Both suspensions and monolayer cultures of the indicator cells can be used. Neutralization of a fraction of a high-titer virus preparation can be scored after the first replication cycle at 8 to 10 h. By lowering the incubation temperature to 30 degrees C, the completion of the cytolysis due to the first replication cycle of poliovirus was delayed beyond 21 h. This makes it possible to use the RACINA, unlike the standard microneutralization assay, for measuring antibodies to trypsin-cleaved polioviruses. The RACINA was found to be as sensitive as and more reproducible than the standard microneutralization assay in the measurement of neutralizing poliovirus antibodies. The RACINA is a rapid and reliable test for neutralizing antibodies and in principle it may be applicable for quantitation of neutralizing antibodies to other cytolytic agents as well.
我们开发了一种新的快速检测方法，即放射细胞溶解抑制试验（RACINA），用于测定脊髓灰质炎病毒中和抗体。预先用51Cr、[3H]亮氨酸，或者优先用[3H]尿苷标记的HeLa细胞用作残留感染性病毒的敏感定量指标。指示细胞的悬浮液和单层培养物均可使用。在8至10小时的第一个复制周期后，即可对高滴度病毒制剂的一部分中和情况进行评分。通过将孵育温度降至30℃，脊髓灰质炎病毒第一个复制周期导致的细胞溶解完成时间延迟至21小时以上。这使得RACINA与标准微量中和试验不同，可用于检测针对胰蛋白酶裂解脊髓灰质炎病毒的抗体。在检测脊髓灰质炎病毒中和抗体时，发现RACINA与标准微量中和试验一样灵敏，且重复性更好。RACINA是一种快速可靠的中和抗体检测方法，原则上也可用于定量其他溶细胞剂的中和抗体。
