An extensive polymerase chain reaction-allele-specific polymorphism strategy for clinical ABO blood group genotyping that avoids potential errors caused by null, subgroup, and hybrid alleles
An extensive polymerase chain reaction-allele-specific polymorphism strategy for clinical ABO blood group genotyping that avoids potential errors caused by null, subgroup, and hybrid alleles
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DOI：
10.1111/j.1537-2995.2007.01436.x
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发表时间：
2007-11-01
期刊：
影响因子：
2.9
通讯作者：
Olsson, Martin L.
中科院分区：
文献类型：
--
作者：
Hosseini-Maaf, Bahram;Hellberg, Asa;Olsson, Martin L.
背景：ABO基因分型由于ABO位点的显著多样性而变得复杂。共同等位基因之间的重组或基因转换可能导致杂种产生意想不到的ABO表型。此外，应该考虑与弱亚群和非缺失空等位基因相关的许多突变。所有已知的ABO基因分型方法，然而，风险不正确的表型预测，如果任何这样的等位基因存在。研究设计和方法：设计了一套广泛的等位基因特异性引物，以完成杂交防多重聚合酶链反应（PCR）扩增DNA片段，用于检测ABO等位基因。结果与血清学结果和先前发表的pcr -限制性片段长度多态性/ pcr -等位基因特异性多态性（ASP）方法或DNA测序确定的ABO基因型进行了比较。结果：对来自普通血型和罕见亚组家族的献血者的表型特征良好的样本进行了分析。除了常见的等位基因(A(1)， A(1（467C > T))， A(2)， B, O-1， O-1v和O-2）外，由于在内含子6上的长距离扩增，新方法可以检测到杂交等位基因。12个PCR-ASP程序中有4个用于筛选多个罕见亚群和空等位基因。这个概念允许使用低分辨率的类型格式，在这种格式中，例如，存在弱子组或cis-AB/B(a)，但没有进一步定义。在可选的高分辨率步骤中，可以获得更详细的基因型信息。结论：一种新的基因分型方法已经被开发和评估，可以正确地识别ABO等位基因，包括非缺失的零等位基因、亚群和由6和7外显子之间的重组交叉事件产生的杂交基因。与以前发表的方法相比，这种方法在临床上是适用的，并且降低了错误ABO表型预测的风险。
Background: ABO genotyping is complicated by the remarkable diversity at the ABO locus. Recombination or gene conversion between common alleles may lead to hybrids resulting in unexpected ABO phenotypes. Furthermore, numerous mutations associated with weak subgroups and nondeletional null alleles should be considered. All known ABO genotyping methods, however, risk incorrect phenotype predictions if any such alleles are present.Study Design and Methods: An extensive set of allele-specific primers was designed to accomplish hybrid-proof multiplex polymerase chain reaction (PCR) amplification of DNA fragments for detection of ABO alleles. Results were compared with serologic findings and ABO genotypes defined by previously published PCR-restriction fragment length polymorphism/PCR-allele-specific polymorphism (ASP) methods or DNA sequencing.Results: Phenotypically well-characterized samples from blood donors with common blood groups and rare-subgroup families were analyzed. In addition to the commonly encountered alleles (A(1), A(1(467C > T)), A(2), B, O-1, O-1v, and O-2), the new method can detect hybrid alleles thanks to long-range amplification across intron 6. Four of 12 PCR-ASP procedures are used to screen for multiple infrequent subgroup and null alleles. This concept allows for a low-resolution typing format in which the presence of, for example, a weak subgroup or cis-AB/B(A) is indicated but not further defined. In an optional high-resolution step, more detailed genotype information is obtained.Conclusion: A new genotyping approach has been developed and evaluated that can correctly identify ABO alleles including nondeletional null alleles, subgroups, and hybrids resulting from recombinational crossing-over events between exons 6 and 7. This approach is clinically applicable and decreases the risk for erroneous ABO phenotype prediction compared to previously published methods.