Meyer Stefan, Vollmert Caren, Trost Nadine, Brönnimann Chantal, Gottschalk Jochen, Buser Andreas, Frey Beat M, Gassner Christoph
Department of Molecular Diagnostics & Cytometry (MOC), Swiss Red Cross, Schlieren, Switzerland.
Transfusion. 2014 Dec;54(12):3198-207. doi: 10.1111/trf.12715. Epub 2014 May 21.
After the ABO (ABO) and Rh (RHD and RHCE) blood group systems, Kell (KEL), Kidd (SLC14A1), and Duffy (DARC) represent the second most important clinically relevant antigens.
Samples from 4000 Swiss blood donors, with serologic prevalues for K/k, Kp(a/b), Jk(a/b), and Fy(a/b), and 48 additional samples of presumptive black African origin were genotyped using high-throughput matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry, applying one single-multiplex polymerase chain reaction/primer-extension reaction simultaneously detecting 15 single-nucleotide polymorphisms.
Genotype/phenotype concordance for K/k, Kp(a/b), Jk(a/b), and all Fy(a/b) specificities were 100, 99.98, 99.93, and 99.20%, respectively. Discrepancies were caused by erroneous serologic profiles (n = 33), mainly attributed to weakly expressed Fy(x) (n = 28). Only three discrepancies had a genetic basis. They could all be explained by newly observed silenced alleles: one KEL02N.34 and one FY02N.03 with predicted R700Q and G261R amino acid exchanges, respectively, and one JKB, with an as-yet-unidentified silencing cause. According to NCBI SNP database entry for rs8176034, another new allele, KEL02.38, had been expected, and we formally demonstrated its presence. We furthermore identified individuals with rare phenotypes, such as Js(a/b) heterozygotes among Caucasians, rare alleles, the "Swiss" JK*01N.03, and rare genotypes, such as one Fy(x) homozygote.
Genotyping proved its practicability in the daily routine setting and qualitatively outperformed serology. Technology is ideal for time-insensitive donor genotyping and allows for a broad range of throughput needs. Consequently, from a technologic point of view, serotyping should be replaced by genotyping for donors' blood groups encoded by KEL, SLC14A1, and DARC.
在ABO（ABO）和Rh（RHD和RHCE）血型系统之后，凯尔（KEL）、基德（SLC14A1）和达菲（DARC）代表了临床上第二重要的相关抗原。
对4000名瑞士献血者的样本进行基因分型，这些样本具有K/k、Kp(a/b)、Jk(a/b)和Fy(a/b)的血清学预值，另外对48份推测为非洲黑人血统的样本也进行了基因分型，采用高通量基质辅助激光解吸/电离飞行时间质谱法，通过一个单重多重聚合酶链反应/引物延伸反应同时检测15个单核苷酸多态性。
K/k、Kp(a/b)、Jk(a/b)以及所有Fy(a/b)特异性的基因型/表型一致性分别为100%、99.98%、99.93%和99.20%。差异是由错误的血清学图谱（n = 33）引起的，主要归因于Fy(x)表达较弱（n = 28）。只有三个差异有遗传基础。它们都可以用新观察到的沉默等位基因来解释：一个KEL02N.34和一个FY02N.03，分别预测有R700Q和G261R氨基酸交换，还有一个JKB，其沉默原因尚未确定。根据NCBI SNP数据库中rs8176034的条目，预计会有另一个新等位基因KEL02.38，我们正式证明了它的存在。我们还鉴定出了具有罕见表型的个体，如白种人中的Js(a/b)杂合子、罕见等位基因“瑞士”JK*01N.03，以及罕见基因型，如一个Fy(x)纯合子。
基因分型在日常工作中证明了其实用性，并且在质量上优于血清学检测。该技术非常适合对时间要求不高的献血者基因分型，并且能够满足广泛的通量需求。因此，从技术角度来看，对于由KEL、SLC14A1和DARC编码的献血者血型，血清分型应该被基因分型所取代。
