风疹病毒IgG抗体参考品的建立

Establishment of Rubella virus IgG antibody reference materials

目的:建立人风疹病毒IgG抗体参考盘,为风疹病毒IgG抗体检测试剂盒的性能评价提供可靠的质控物质。方法:通过风疹病毒IgG抗体检测试剂盒进行血清筛选和复核,免疫荧光方法和Western blot方法对筛选的血清进行确认,获得了符合要求的血清,组成了含有10份阳性血清和5份阴性血制成参考品的P1～P10和N1～N5、重复性参考品R以及检测限参考品L1,共有17支参考品的参考盘。经过不同方法学的性能验证,确立风疹病毒IgG抗体参考盘的性能指标,并且参考盘的稳定性进行测试。结果:经风疹病毒IgG抗体检测试剂盒初筛和复核、以及免疫荧光法和Western blot方法确认,确定10份风疹病毒IgG阳性血清样本和5份阴性血清样本,选择其中一份阳性样本作为重复性参考品,10 IU.mL-1作为检测限参考品,构建成人风疹病毒IgG抗体参考盘。确定了参考盘的性能指标:1.准确性(阳性符合率):检测国家阳性参考品10份样本,要求假阴性应不得出现。2.特异性(阴性符合率):检测国家阴性参考品5份样本,要求假阳性应不得出现。3.重复性(精密度):批内精密度RSO不大于15.0%。4.检测限:用风疹病毒IgG抗体检测试剂对国家最低检出限参考品进行检测,要求L1应能检出。稳定性研究结果表明,参考盘的稳定性良好。成功地建立了人风疹病毒IgG抗体国家参考盘。结论:该参考盘为临床实验室质量评价提供了可靠的参考物质,可用于国内大多数试剂盒的性能评价要求。

Objective： To establish the reference materials of human Rubella virus IgG antibody, providing the relia- ble quality control materials for the performance evaluation of Rubella virus IgG antibody detection kit. Method：Se- ra were screened and reviewed by Rubella virus IgG antibody detection kit. Then the sera accorded with require- ments were obtained and confirmed by immunofluorescence assay and western blot assay. The obtained sera constitu- ted 17 reference materials, including positive reference materials （ P1 - P10 ）, negative reference materials （ N1 - N5 ）, repetitive reference material（R） and detection limit reference material（ L1 ）. Performance indexes of reference materials were established through various performance verifications and the stability of reference materials was in- vestigated at different times under different conditions. Results：Through the preliminary screening and reviewing by Rubella virus IgG antibody detection kit and confirming Rubella virus IgG antibody positive samples and 5 negative by immunofluorescence assay and western blot assay, 10 samples were confirmed, of which one positive sample was taken as the repeated reference, constituting the human Rubella virus IgG antibody reference plate with 10 IU · mL- 1 as the detection limit. The performance indexes for reference materials were established： 1. Accuracy （positive coincidence rate） ：the detection of 10 positive reference materials should have no false negative. 2. Specificity（ nega- tive coincidence rate） ：the detection of 5 negative reference materials should have no false positive. 3. Reproducibili- ty（precision） ：RSD of precision was no more than 15.0%. 4. Detection limit：the detection limit reference material should be positive. The results of stability showed the stability of reference materials were fine. Therefore, the nation- al reference materials of human Rubella virus IgG antibody were established. Conclusion：The reference materials provide the reliable reference materials for quality evaluation in clinical laboratory, which can be used for the per-formance evaluation of most detection kits.