Glycophorins are the most abundant sialoglycoproteins on the surface of human erythrocyte membranes. Genetic variation in glycophorin region of human chromosome 4 (containing GYPA, GYPB, and GYPE genes) is of interest because the gene products serve as receptors for pathogens of major public health interest, including Plasmodium sp., Babesia sp., Influenza virus, Vibrio cholerae El Tor Hemolysin, and Escherichia coli. A large structural rearrangement and hybrid glycophorin variant, known as Dantu, which was identified in East African populations, has been linked with a 40% reduction in risk for severe malaria. Apart from Dantu, other large structural variants exist, with the most common being deletion of the whole GYPB gene and its surrounding region, resulting in multiple different deletion forms. In West Africa particularly, these deletions are estimated to account for between 5 and 15% of the variation in different populations, mostly attributed to the forms known as DEL1 and DEL2. Due to the lack of specific variant assays, little is known of the distribution of these variants. Here, we report a modification of a previous GYPB DEL1 assay and the development of a novel GYPB DEL2 assay as high-throughput PCR-RFLP assays, as well as the identification of the crossover/breakpoint for GYPB DEL2. Using 393 samples from three study sites in Ghana as well as samples from HapMap and 1000 G projects for validation, we show that our assays are sensitive and reliable for genotyping GYPB DEL1 and DEL2. To the best of our knowledge, this is the first report of such high-throughput genotyping assays by PCR-RFLP for identifying specific GYPB deletion types in populations. These assays will enable better identification of GYPB deletions for large genetic association studies and functional experiments to understand the role of this gene cluster region in susceptibility to malaria and other diseases.

糖蛋白是人红细胞膜表面最丰富的唾液酸糖蛋白。人类 4 号染色体的血型糖蛋白区域（包含GYPA、GYPB和GYPE基因）的遗传变异是令人感兴趣的，因为基因产物可作为主要公共卫生利益病原体的受体，包括疟原虫 。，巴贝虫 属。、流感病毒、霍乱弧菌El Tor 溶血素和大肠杆菌。一种大的结构重排和混合血型糖蛋白变体，称为Dantu在东非人群中发现的这种疾病与严重疟疾的风险降低 40% 有关。除丹兔外，还存在其他较大的结构变异，最常见的是整个GYPB基因及其周围区域的缺失，导致多种不同的缺失形式。特别是在西非，这些缺失估计占不同人群变异的 5% 到 15%，主要归因于称为 DEL1 和 DEL2 的形式。由于缺乏特定的变体测定，对这些变体的分布知之甚少。在这里，我们报告了对先前GYPB DEL1 检测的修改和新型GYPB的开发DEL2 测定作为高通量 PCR-RFLP 测定，以及GYPB DEL2 交叉/断点的鉴定。使用来自加纳三个研究地点的 393 个样本以及来自 HapMap 和 1000 G 项目的样本进行验证，我们表明我们的分析对GYPB DEL1 和 DEL2 的基因分型是敏感和可靠的。据我们所知，这是通过 PCR-RFLP 进行这种高通量基因分型测定的第一份报告，用于鉴定群体中特定的GYPB缺失类型。这些化验将能够更好地识别 GYPB 缺失，用于大型遗传关联研究和功能实验，以了解该基因簇区域在疟疾和其他疾病易感性中的作用。