van der Vlies S A, Voorter C E, van den Berg-Loonen E M
Tissue Typing Laboratory, University Hospital Maastricht, The Netherlands.
Tissue Antigens. 1998 Dec;52(6):558-68. doi: 10.1111/j.1399-0039.1998.tb03087.x.
Serological typing of HLA-C has been poor and almost half of its alleles are serologically undetectable blanks in most populations. Therefore, DNA typing techniques have been used to identify and type for the HLA-C gene. Sequence-based typing (SBT) has proven a major typing strategy for highly polymorphic HLA genes. The technique enables direct identification of all sequence motifs without the need to continuously adjust primers. Here we describe a reliable solid-phase SBT strategy for HLA-C which can be used to distinguish all currently known HLA-C alleles without prior knowledge gained by low resolution typing. Exons 2 and 3 were amplified and sequenced and if necessary sequences of exons 1 and 5 were determined. A total of 257 individuals were typed for HLA-C using this protocol and 30 of the 42 known HLA-C alleles were detected. All heterozygous combinations found in this study were unambiguously discriminated. One hundred and forty-four individuals from the Dutch population were typed randomly. In this group Cw0701 and 0702 were the most frequently detected alleles. Of the serological Cw blank alleles Cw1203 was found to have the highest frequency (16%). From the total group 212 individuals were typed serologically and 106 were retyped with 97 selected antisera to further compare serological and molecular defined phenotypes. Discrepancies between serological typing and SBT are mainly attributable to the serologically Cw blank alleles Cw12-18. The high resolution SBT protocol described will be a valuable tool for the identification of HLA-C alleles and the determination of the role of HLA-C in marrow and organ transplantation.
HLA - C的血清学分型效果不佳，在大多数人群中，其几乎一半的等位基因在血清学上无法检测到，表现为空白。因此，DNA分型技术已被用于HLA - C基因的鉴定和分型。基于序列的分型（SBT）已被证明是高度多态性HLA基因的主要分型策略。该技术能够直接识别所有序列基序，无需不断调整引物。在此，我们描述了一种可靠的用于HLA - C的固相SBT策略，该策略可用于区分所有目前已知的HLA - C等位基因，而无需通过低分辨率分型预先获得相关知识。对第2和第3外显子进行扩增和测序，如有必要，确定第1和第5外显子的序列。使用该方案对总共257名个体进行了HLA - C分型，检测到了42个已知HLA - C等位基因中的30个。在本研究中发现的所有杂合组合都得到了明确区分。随机对144名荷兰人群个体进行了分型。在该组中，Cw0701和0702是最常检测到的等位基因。在血清学Cw空白等位基因中，发现Cw1203的频率最高（16%）。从整个组中，对212名个体进行了血清学分型，并用97种选定的抗血清对其中106名个体进行了重新分型，以进一步比较血清学和分子定义的表型。血清学分型与SBT之间的差异主要归因于血清学Cw空白等位基因Cw12 - 18。所描述的高分辨率SBT方案将成为鉴定HLA - C等位基因以及确定HLA - C在骨髓和器官移植中作用的宝贵工具。
