RAPID DNA TYPING FOR HLA-C USING SEQUENCE-SPECIFIC PRIMERS (PCR-SSP) - IDENTIFICATION OF SEROLOGICAL AND NON-SEROLOGICALLY DEFINED HLA-C ALLELES INCLUDING SEVERAL NEW ALLELES
RAPID DNA TYPING FOR HLA-C USING SEQUENCE-SPECIFIC PRIMERS (PCR-SSP) - IDENTIFICATION OF SEROLOGICAL AND NON-SEROLOGICALLY DEFINED HLA-C ALLELES INCLUDING SEVERAL NEW ALLELES
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DOI：
10.1111/j.1399-0039.1994.tb02290.x
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发表时间：
1994-01-01
期刊：
影响因子：
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通讯作者：
WELSH, KI
中科院分区：
文献类型：
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作者：
BUNCE, M;WELSH, KI
用人类同种异体血清通过补体介导的细胞毒作用来检测人类白细胞抗原-C往往是困难的。在每个种族中，有20%~40%的个体具有无法检测到的HLA-C位点抗原，目前已发表的29个已测序的HLA-C等位基因中有9个编码了血清学上未检测到的抗原。此外，人类白细胞抗原-C分子在细胞表面的表达水平约为人类白细胞抗原A和B的10%。近年来，DNA序列特异性扩增技术(PCR-SSP)已被证明是一种可靠、快速的检测人类白细胞抗原DR、DQA和DQB基因的方法。PCR-SSP需要两个小时的时间，适合身体捐献者的基因分型。我们设计了一套能正确识别出与血清学上定义的系列-Cw1、Cw2、Cw3、Cw4、Cw5、Cw6、Cw7和Cw8相对应的HLA-C等位基因的引物。根据序列同源性，还检测到血清学上无法检测到的等位基因。此外，还发现了三个新的未测序变异体。对56个国际组织相容性研讨会参考细胞系和103个对照个体的DNA样本进行了人类白细胞抗原-C聚合酶链式反应-单链构象多态分析。4/56细胞系类型和11/103正常对照类型与报道的血清学类型不一致。可以很容易地识别出血清学上可检测到的所有组合和大多数血清学上空白的HLA-C抗原。用聚合酶链式反应-单链构象多态技术对人类白细胞抗原Cw进行DNA分型，从开始到结束只需要130分钟，包括DNA准备。
Detection of HLA-C antigens by complement mediated cytotoxicity using human alloantisera is often difficult. Between 20 to 40% of individuals in every race have undectectable HLA-C locus antigens and 9 out of the 29 sequenced HLA-C alleles so far published encode serologically undetected antigens. In addition, HLA-C molecules are expressed at the cell surface at about 10% of the levels of HLA-A and HLA-B. Recently, amplification of DNA using sequence-specific primers (PCR-SSP) has proved a reliable and rapid method for typing HLA-DR, HLA-DQA and HLA-DQB genes. PCR-SSP takes two hours to perform and is therefore suitable for the genotyping of cadaveric donors. We have designed a set of primers which will positively identify the HLA-C alleles corresponding to the serologically defined series HLA-Cw1, Cw2, Cw3, Cw4, Cw5, Cw6, Cw7 and Cw8. The serologically undetectable alleles have also been detected in groups according to sequence homology. In addition, three new unsequenced variants have been identified. DNA samples from 56 International Histocompatibility Workshop reference cell lines and 103 control individuals have been typed by the HLA-C PCR-SSP technique. 4/56 cell line types and 11/103 normal control individuals types were discrepant with the reported serological types. Ail combinations of serologically detectable and most of the serologically blank HLA-C antigens can be readily identified. DNA typing for HLA-Cw by PCR-SSP can take as little as 130 minutes from start to finish, including DNA preparation.