Yu N, Ohashi M, Alosco S, Granja C, Salazar M, Hegland J, Yunis E
American Red Cross Blood Services, New England Region, Dedham, Massachusetts 02026, USA.
Tissue Antigens. 1997 Oct;50(4):380-6. doi: 10.1111/j.1399-0039.1997.tb02891.x.
We are reporting the results of HLA-A typing by PCR-SSOP complemented by PCR-SSP of samples obtained from the National Marrow Donor Program (NMDP). These samples were a representative group from 2486 tested in duplicate by serology. A total of 390 samples gave HLA-A discrepant results. Comparing the molecular typing results of 238 samples (samples with available DNA) with the serological typing results, 54 homozygotes and 184 heterozygotes produced a total of 422 assignments by molecular methods. We found assignment discrepancies in 147/422 (35%) in laboratory 1 and 144/422 (34%) in laboratory 2 (a combined group of 4 NMDP laboratories; laboratory 1 is not included). The serological discrepancies found were of 3 categories: a) false negatives, b) incomplete typing (discrepancies due to the level of resolution within a cross-reactive or CREG group) and c) false positives. Major problems were identified using serology for typing HLA-A antigens: a) inability to identify all WHO-recognized specificities, more frequently in non-Caucasians or in HLA-A specificities known to be found more frequently in non-Caucasians for laboratory 1 and incorrect assignments of A19 specificities in laboratory 2, b) incorrect assignments in cells with poor viability and c) false-positive assignments in homozygotes. We propose a possible strategy to type HLA-A specificities with two steps: a) a minimum of serology for typing specificities for common CREG groups: A1, A2, A3, A11, A9, A10, A28, A19. However, a given laboratory can determine the level of serological assignments needed as a first step. And b) molecular methods to identify splits: A23, A24, A29, A30, A31, A32, A33, A34, A36, A66, A74 and A80. The technique described is useful for large-scale bone marrow donor typings for cells with poor viability, and for resolving ambiguous results including false-positive assignments of homozygous cells.
我们报告了通过聚合酶链反应-序列特异性寡核苷酸探针杂交（PCR-SSOP）并辅以聚合酶链反应-序列特异性引物（PCR-SSP）对从国家骨髓捐赠计划（NMDP）获得的样本进行HLA-A分型的结果。这些样本是从2486份经血清学重复检测的样本中选取的具有代表性的一组。共有390份样本的HLA-A分型结果存在差异。将238份样本（有可用DNA的样本）的分子分型结果与血清学分型结果进行比较，54份纯合子和184份杂合子通过分子方法共产生了422个分型结果。我们发现，在实验室1中，422个分型结果中有147个（35%）存在分型差异，在实验室2中（4个NMDP实验室的联合组；不包括实验室1）有144个（34%）存在分型差异。发现的血清学差异有3类：a）假阴性，b）分型不完整（由于交叉反应或CREG组内分辨率水平导致的差异）和c）假阳性。使用血清学进行HLA-A抗原分型时发现了主要问题：a）无法识别所有世界卫生组织认可的特异性，在非白种人中更常见，或者在实验室1中，在已知在非白种人中更常见的HLA-A特异性中，以及在实验室2中A19特异性的错误分型；b）对活力差的细胞进行错误分型；c）纯合子中的假阳性分型。我们提出了一种对HLA-A特异性进行分型的可能策略，分两步进行：a）对常见CREG组的特异性进行分型时，至少进行血清学检测：A1、A2、A3、A11、A9、A10、A28、A19。然而，特定实验室可以确定第一步所需的血清学分型水平。b）使用分子方法识别细分特异性：A23、A24、A29、A30、A31、A32、A33、A34、A36、A66、A74和A80。所描述的技术对于对活力差的细胞进行大规模骨髓供体分型以及解决包括纯合细胞假阳性分型在内的模糊结果很有用。
