HLA class II typing by digestion of PCR-amplified DNA with allele-specific restriction endonucleases will fail to unequivocally identify the genotypes of many homozygous and heterozygous individuals.
HLA class II typing by digestion of PCR-amplified DNA with allele-specific restriction endonucleases will fail to unequivocally identify the genotypes of many homozygous and heterozygous individuals.
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通过用等位基因特异性限制性内切酶消化 PCR 扩增的 DNA 来进行 HLA II 类分型将无法明确鉴定许多纯合和杂合个体的基因型。
DOI：
10.1111/j.1399-0039.1990.tb01805.x
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发表时间：
1990
期刊：
影响因子：
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通讯作者：
O. Olerup
中科院分区：
文献类型：
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作者：
O. Olerup
最近，引入了一种新的HLA II类基因分型技术，即聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法，该方法被认为是常规血清学和细胞HLA II类分型的实用替代方法（1-3）。PCR-RFLP技术是巧妙的，相对快速的，不需要与序列特异性寡核苷酸探针杂交。然而，分析各种研究的HLA II类基因座的PCR-RFLP模式的纯合和杂合组合是否是唯一的，不幸的是，仅19%的DRB纯合和杂合组合是唯一的。DQA 1、DQB和DPB位点的检出率分别为56%、29%和65%。由于并非在上述研究（1-3）中分析的所有核苷酸序列都产生独特的PCR-RFLP，并且由于现在已知更多的序列，对于DRB 1、DRB 3、DRB 5和DPB 1基因座（4），不同HLA II类基因座的独特PCR-RFLP模式的频率将进一步降低。因此，目前的分析表明，PCR-RFLP技术，如参考文献1-3中所述，尚未准备好用于常规HLA II类基因分型。PCR-RFLP方法的分辨率可以通过各种修改来提高。然而，改良的PCR-RFLP技术在HLA II类分型中的作用仍有待证实。
Recently, a new technique for HLA class II genotyping has been introduced, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, claimed to be a practical alternative to conventional serological and cellular HLA class II typing (1-3). The PCR-RFLP technique is ingenious, relatively rapid and does not require hybridization with sequence-specific oligonucleotide probes. However, analysis of whether homozygous and heterozygous combinations of PCR-RFLP patterns for the various investigated HLA class II loci are unique or not unfortunately shows that only 19% of DRB homozygous and heterozygous combinations are unique. The figures for the DQA1, DQB and DPB loci are 56%, 29% and 65%, respectively. As not all nucleotide sequences analyzed in the above-mentioned studies (1-3) gave rise to unique PCR-RFLPs and as more sequences now are known, for the DRB1, DRB3, DRB5 and DPB1 loci (4), the frequencies of unique PCR-RFLP patterns for the different HLA class II loci will be reduced even further. Thus, the present analysis demonstrates that the PCR-RFLP technique, performed as described in references 1-3, is not yet ready to be used for routine HLA class II genotyping. The resolution of the PCR-RFLP method can be improved by various modifications. However, the role of a modified PCR-RFLP technique in HLA class II typing still remains to be shown.