A flexible array format for large-scale, rapid blood group DNA typing
A flexible array format for large-scale, rapid blood group DNA typing
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DOI：
10.1111/j.1537-2995.2005.04362.x
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发表时间：
2005-05-01
期刊：
影响因子：
2.9
通讯作者：
Reid, ME
中科院分区：
文献类型：
--
作者：
Hashmi, G;Shariff, T;Reid, ME
背景：血型抗原的分型目前是通过血凝进行的。必要的试剂变得昂贵且可用性有限，并且方法是劳动密集型的。本研究的目的是确定使用大规模的DNA分析在微阵列作为一种替代血型typing.Study设计和方法的可行性：DNA，提取的血液样本已被表型的一些红细胞抗原，分析了选定的血型等位基因珠阵列（BeadChip，Illumina）和手动聚合酶链反应（PCR）为基础的测定。通过酶介导的探针延伸来鉴定选定的等位基因，所述探针位于在硅芯片上组装成阵列的颜色编码珠上。使用来自血清学表征的供体的DNA验证了包含FYA/B、FY-GATA、DOA/B、COA/B、LWA/B、DIA/B和SC 1/SC 2单核苷酸多态性（SNP）的原型BeadChip（BLOOD-1）的性能。然后，它被用来分析400多个部分定义的表型样本。用BLOOD-1检测来自中国、德系和泰国供体的样本（总计n = 227）。一个扩展的珠芯片，共有18个SNP（36个等位基因;然后用来自中国人、德系犹太人、犹太人、犹太人和犹太人的先前测试样品的子集评估血液-1和M/N、S/s、Lu-a/Lu-B、K/k、FY 265 [用于Fy（X）多态性]、Jk（a）/Jk（B）、DO 323 [用于Hy]、DO 350 [用于Jo（a）]和HgbS）中的SNP。结果：BeadChip检测结果与手工PCR-限制性片段长度多态性、等位基因特异性PCR和血凝试验结果一致。来自不同种族的样本中的等位基因的频率在预期的范围内，但是，两个新的DO等位基因被发现。结论：它已被证明，基因芯片技术可以用于类型的DNA和检测新的等位基因在供体队列。
BACKGROUND: Typing for blood group antigens is currently performed by hemagglutination. The necessary reagents are becoming costly and limited in availability, and the methods are labor-intensive. The purpose of this study was to determine the feasibility of the use of large-scale DNA analysis in a microarray as a substitute for blood group typing.STUDY DESIGN AND METHODS: DNA, extracted from blood samples that had been phenotyped for some of the red blood cell antigens, was analyzed for selected blood group alleles by bead array (BeadChip, Illumina) and by manual polymerase chain reaction (PCR)-based assays. Selected alleles were identified by enzyme-mediated elongation of probes, which were on color-encoded beads assembled into arrays on silicon chips. The performance of a prototype BeadChip (BLOOD-1) containing single-nucleotide polymorphisms (SNPs) for FYA/B, FY-GATA, DOA/B, COA/B, LWA/B, DIA/B, and SC1/SC2 was verified with DNA from serologically characterized donors. It was then used to analyze more than 400 samples of partially defined phenotype. Samples from Chinese, Ashkenazi, and Thai donors (total n = 227) were tested with BLOOD-1. An expanded BeadChip with a total of 18 SNPs (36 alleles; SNPs in BLOOD-1 and M/N, S/s, Lu-a/Lu-b, K/k, FY265[for the Fy(X) polymorphism], Jk(a)/Jk(b), DO323[for Hy], DO350[for Jo(a)], and HgbS) was then evaluated with a subset of previously tested samples from Chinese, Ashkenazi, and New York blood donors (127) and an additional set of samples from Israeli donors (total n = 188).RESULTS: Results obtained by BeadChip analysis were concordant with those obtained with the manual PCR-restriction fragment length polymorphism, allele-specific PCR, and hemagglutination assays. The frequencies of the alleles in the samples from different ethnic panels were within the expected ranges; however, two new DO alleles were discovered.CONCLUSION: It has been shown that microarray technology can be used to type DNA and detect new alleles in donor cohorts.