POLYMORPHISM OF HLA-DRW52-ASSOCIATED DRB1 GENES AS DEFINED BY SEQUENCE-SPECIFIC OLIGONUCLEOTIDE PROBE HYBRIDIZATION AND SEQUENCING
POLYMORPHISM OF HLA-DRW52-ASSOCIATED DRB1 GENES AS DEFINED BY SEQUENCE-SPECIFIC OLIGONUCLEOTIDE PROBE HYBRIDIZATION AND SEQUENCING
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DOI：
10.1111/j.1399-0039.1991.tb01891.x
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发表时间：
1991-10-01
期刊：
影响因子：
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通讯作者：
HANSEN, JA
中科院分区：
文献类型：
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作者：
PETERSDORF, EW;SMITH, AG;HANSEN, JA
我们利用群体特异性DNA扩增和序列特异性寡核苷酸探针（SSOP）杂交研究了DRB1序列与DR3、DRw11(5)、DRw12(5)、DRw13（w6）、DRw14（w6）和DRw8等位基因相关的多态性。使用设计用于与该组所有已知等位基因共同的第一高变区（HVR）序列杂交的5‘扩增引物，以及3’内含子引物，实现了drw52相关DRB1等位基因的群体特异性扩增。对318例患者进行DR3、DRw11、DRw12、DRw13、DRw14和DRw8等位基因的SSOP分型，包括124例患者、46名家庭成员和148名无血缘关系的骨髓供者。在本研究检测到的395个drw52相关DRB1等位基因中，除6例（1.9%）外，所有个体均能找到与先前定义的等位基因DRB1*0301-2 （DR3）、DRB1*1101-4 （DR5）、DRB1*1201-2 （DR5）、DRB1*1301-5 （DRw6）、DRB1*1401-2和1404 （DRw6）以及DRB1*0801-4 （DRw8）相对应的亚型。除了已知的22个等位基因外，我们还鉴定了两个新的drw6相关等位基因。DRB1 * 13。MW(1)和DRB1*14.GB(1)。DRB1 * 13。MW的血清学分型为DRw13，与DRB1*1301相同，但密码子71处AGG编码精氨酸，而GAG编码谷氨酸。DRB1 * 14。GB表示DRB1*1402变异，其密码子86处的序列编码缬氨酸（GTG）而不是甘氨酸（GGT）。这些结果表明，SSOP方法是一种高效、精确的DRB1等位基因分型方法，并可用于识别传统分型方法无法识别的潜在新变异。
We have used group-specific DNA amplification and sequence-specific oligonucleotide probe (SSOP) hybridization to study DRB1 sequence polymorphisms associated with DR3, DRw11(5), DRw12(5), DRw13(w6), DRw14(w6) and DRw8 alleles. Group-specific amplification of DRw52-associated DRB1 alleles was achieved using a 5' amplification primer designed to hybridize with a first hypervariable region (HVR) sequence common to all known alleles in this group, together with a 3' intron primer. Prospective SSOP typing of DR3, DRw11, DRw12, DRw13, DRw14 and DRw8 alleles was performed in 318 individuals, including 124 patients, 46 family members and 148 unrelated marrow donors. Among the 395 DRw52-associated DRB1 alleles tested in our study, a subtype corresponding to the previously defined alleles DRB1*0301-2 (DR3), DRB1*1101-4 (DR5), DRB1*1201-2 (DR5), DRB1*1301-5 (DRw6), DRB1*1401-2 and 1404 (DRw6), and DRB1*0801-4 (DRw8) could be assigned in all but 6 individuals (1.9%) tested. In addition to the 22 known alleles, we identified two new DRw6-associated alleles. DRB1*13.MW(1) and DRB1*14.GB(1). DRB1*13.MW typed serologically as DRw13 and was identical to DRB1*1301 except at codon 71 where AGG encodes arginine instead of GAG encoding glutamic acid. DRB1*14.GB represents a DRB1*1402 variant whose sequence at codon 86 encodes valine (GTG) instead of glycine (GGT). These results demonstrate that SSOP methods represent an efficient and precise approach for typing DRB1 alleles and for identifying potential novel variants previously unrecognized by conventional typing methods.