Verduyn W, Anholts J D, Versluis L F, Parlevliet J, Drabbels J, De Meester J, Tilanus M G, Doxiadis I I, Giphart M J, Schreuder G M
Department of Immunohematology and Bloodbank, Leiden University Hospital, The Netherlands.
Tissue Antigens. 1996 Aug;48(2):80-6. doi: 10.1111/j.1399-0039.1996.tb02611.x.
Seven samples with irregular PCR-SSO hybridization patterns, observed during routine HLA-DRB typing, were studied in more detail. Group-specific amplification, followed by hybridization with relevant SSOs strengthened the suggestion that these samples contained new DRB alleles. DRB exon 2 segments were amplified, cloned and sequenced and revealed: DRB11121 [MUL] is similar to DRB11102 in which codon 85 changed from GTT(V) into GTC(A); DRB11419 [AKKAL] is similar to DRB11402 with codon 71 changed from AGG(R) into AAG(K); DRB11420 [OND-52971] is a DRB11406 with codon 37 changed from AAC(N) into TTC(F); DRB11421 [TGI] is similar to DRB11417 with codon 71 changed from AGG(R) into AAG(K); DRB30203 [POS] is similar to DRB30202 in which codons 37-38 are changed from TAC GCG(YA) into TCC GTC(SV); DRB50103 was found in two unrelated individuals of Oriental origin [IND-24 and IND-59] and is similar to DRB50102 in which codon 71 AGG(R) changed into ACG(T). This particular sequence variation at position 71 has not yet been described. The new DRB sequences were confirmed using the sequencing based typing technique. Low resolution PCR-SSP typing failed to amplify two of the DRB114 variants, whereas high resolution PCR-SSP resulted in aberrant patterns. Class II alloantisera identify the codon 71 changes in DRB11419 and *1421 with respect to the MC1 ('DR1+DR4') epitope.
在常规HLA - DRB分型过程中观察到7个具有不规则PCR - SSO杂交模式的样本,对其进行了更详细的研究。通过组特异性扩增,随后与相关的SSO进行杂交,进一步表明这些样本含有新的DRB等位基因。对DRB外显子2片段进行扩增、克隆和测序后发现:DRB11121 [MUL]与DRB11102相似,其中第85密码子从GTT(V)变为GTC(A);DRB11419 [AKKAL]与DRB11402相似,第71密码子从AGG(R)变为AAG(K);DRB11420 [OND - 52971]是DRB11406,第37密码子从AAC(N)变为TTC(F);DRB11421 [TGI]与DRB11417相似,第71密码子从AGG(R)变为AAG(K);DRB30203 [POS]与DRB30202相似,其中第37 - 38密码子从TAC GCG(YA)变为TCC GTC(SV);DRB50103在两名无关的东方血统个体[IND - 24和IND - 59]中被发现,与DRB50102相似,其中第71密码子AGG(R)变为ACG(T)。第71位的这种特定序列变异尚未见报道。使用基于测序的分型技术对新的DRB序列进行了确认。低分辨率PCR - SSP分型未能扩增出两个DRB114变体,而高分辨率PCR - SSP则产生异常模式。II类同种抗血清可识别DRB11419和*1421中相对于MC1(“DR1 + DR4”)表位的第71密码子变化。