Dall'Ozzo Sébastien, Andres Christian, Bardos Pierre, Watier Hervé, Thibault Gilles
EA 3249, Group, Laboratoire d'Immunologie, Faculté de Médecine, 2 bis Boulevard Tonnellé, 37032 Cedex, Tours, France.
J Immunol Methods. 2003 Jun 1;277(1-2):185-92. doi: 10.1016/s0022-1759(03)00123-6.
A polymorphism at position 559 in the cDNA of the FCGR3A gene encoding the FcgammaRIIIa generates two allotypes with either a valine (Val) or a phenylalanine (Phe) at amino acid position 158. This polymorphism is of major importance in immunopathology and in pharmacogenetics, especially monoclonal antibody treatments. In this study, we report a single-step and single-tube method for FCGR3A-158V/F genotyping by real-time multiplex allele-specific PCR and melting curve analysis in the presence of SYBR Green I fluorescent dye. Results obtained from 113 samples showed 100% accuracy compared to those obtained with a conventional PCR-based allele-specific restriction assay. Although this method requires expensive equipment, it is inexpensive in terms of consumables. It is also very rapid, reliable and suitable for large-scale analysis.
编码FcγRIIIa的FCGR3A基因cDNA中第559位的多态性产生了两种同种异型，在氨基酸位置158处分别为缬氨酸（Val）或苯丙氨酸（Phe）。这种多态性在免疫病理学和药物遗传学中，尤其是在单克隆抗体治疗中具有重要意义。在本研究中，我们报告了一种单步单管方法，用于在SYBR Green I荧光染料存在的情况下，通过实时多重等位基因特异性PCR和熔解曲线分析对FCGR3A-158V/F进行基因分型。从113个样本中获得的结果与使用传统的基于PCR的等位基因特异性限制性分析获得的结果相比，准确率为100%。虽然该方法需要昂贵的设备，但就消耗品而言成本较低。它也非常快速、可靠，适用于大规模分析。
