Bozón M V, Delgado J C, Selvakumar A, Clavijo O P, Salazar M, Ohashi M, Alosco S M, Russell J, Yu N, Dupont B, Yunis E J
Dana-Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Tissue Antigens. 1997 Oct;50(4):387-94. doi: 10.1111/j.1399-0039.1997.tb02892.x.
Until recently, the majority of HLA class I typing has been performed by serology. Expensive commercial typing trays are frequently used for testing non-Caucasian subjects and new strategies using DNA-based methods have been adopted for improving clinical histocompatibility testing results and adapted as supplements in proficiency testing. A double-blind comparison of the typing of HLA-B specificities in 40 samples was carried out between serology and two polymerase chain reaction (PCR) methods, PCR amplification with sequence-specific primers (PCR-SSP) and PCR amplification and subsequent hybridization with sequence-specific oligonucleotide probes (PCR-SSOP). The results demonstrated 22.5% misassignments of HLA-B antigens by serology. There was complete concordance between the results obtained with the two PCR based typing methods. A second panel of 20 donor samples with incomplete or ambiguous serologic results was analyzed by PCR-SSP and SSOP Both PCR methods identified correctly the HLA-B antigens. Our results suggest that more accurate typing results can be achieved by complementing serologic testing with DNA-based typing techniques. The level of resolution for HLA-B antigen assignment can be obtained by this combination of serology and limited DNA-based typing is equivalent to the HLA-B specificities defined by the WHO-HLA Committee. This level of resolution cannot routinely be achieved in clinical histocompatibility testing or in proficiency testing using serologic reagents only.
直到最近，大多数HLA I类分型仍通过血清学方法进行。昂贵的商业分型板经常用于检测非白种人受试者，并且已采用基于DNA的新方法来改善临床组织相容性检测结果，并将其用作能力验证的补充方法。对40份样本中的HLA - B特异性进行了血清学方法与两种聚合酶链反应（PCR）方法（即序列特异性引物PCR扩增法（PCR - SSP）和PCR扩增及随后与序列特异性寡核苷酸探针杂交法（PCR - SSOP））之间的双盲比较。结果表明，血清学方法对HLA - B抗原的错误分型率为22.5%。两种基于PCR的分型方法所得结果完全一致。对第二组20份血清学结果不完整或不明确的供体样本进行了PCR - SSP和SSOP分析，两种PCR方法均正确鉴定出了HLA - B抗原。我们的结果表明，通过将血清学检测与基于DNA的分型技术相结合，可以获得更准确的分型结果。通过血清学与有限的基于DNA的分型相结合所能达到的HLA - B抗原分型分辨率水平，等同于世界卫生组织HLA委员会所定义的HLA - B特异性。仅使用血清学试剂，在临床组织相容性检测或能力验证中通常无法达到这种分辨率水平。
