Turner D, Akpe S, Brown J, Brown C, McWhinnie A, Madrigal A, Navarrete C
Department of Histocompatibility and Immunogenetics, North London Centre, National Blood Service, London, UK.
Hum Immunol. 2001 Apr;62(4):414-8. doi: 10.1016/s0198-8859(01)00213-0.
The application of reference strand conformation analysis (RSCA) to HLA-A typing using the ABI PRISM 310 capillary based genetic analyzer has recently been described. This study outlines the development and validation of capillary RSCA for HLA-B typing. Mobility values for 93 HLA-B alleles were defined following electrophoresis of known controls through the system. Three fluorescently labelled references, labelled with three different dyes can be electrophoresed simultaneously. The technique was validated by comparing results from 296 cord blood donors with those obtained using reverse SSO. Following capillary RSCA 14.5% of samples required confirmatory typing, compared with a repeat rate of 5.1% following reverse SSO. In samples where no other typing was necessary there was 100% correlation between the two methods. Capillary RSCA for HLA-B typing is quick, easy to implement, and with the introduction of new FLRs and gel matrices has the potential to evolve into a high resolution typing method.
参考链构象分析（RSCA）在使用基于ABI PRISM 310毛细管的基因分析仪进行HLA - A分型中的应用最近已有报道。本研究概述了用于HLA - B分型的毛细管RSCA的开发与验证。通过已知对照在该系统中进行电泳后，确定了93个HLA - B等位基因的迁移率值。三种用不同染料标记的荧光标记参考物可同时进行电泳。通过将296名脐血供者的结果与使用反向序列特异性寡核苷酸（reverse SSO）获得的结果进行比较，对该技术进行了验证。采用毛细管RSCA后，14.5%的样本需要进行确证分型，而采用反向SSO后的重复率为5.1%。在无需进行其他分型的样本中，两种方法的相关性为100%。用于HLA - B分型的毛细管RSCA快速、易于实施，并且随着新的荧光参考物（FLRs）和凝胶基质的引入，有可能发展成为一种高分辨率分型方法。
