Analytical validation of the tag-it high-throughput microsphere-based universal array genotyping platform: Application to the multiplex detection of a panel of thrombophilia-associated single-nucleotide polymorphisms
Analytical validation of the tag-it high-throughput microsphere-based universal array genotyping platform: Application to the multiplex detection of a panel of thrombophilia-associated single-nucleotide polymorphisms
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DOI：
10.1373/clinchem.2004.035071
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发表时间：
2004-11-01
影响因子：
9.3
通讯作者：
Zastawny, R
中科院分区：
文献类型：
--
作者：
Bortolin, S;Black, M;Zastawny, R
背景：我们开发了一种新颖的、基于微球的通用阵列平台，称为Tag-It(TM)平台。该平台适用于高通量的临床基因分型应用，并用于一组血栓相关单核苷酸多态(SNPs)的多重分析。方法：用6组引物对132例患者的基因组DNA进行多重PCR扩增，然后用12个通用标记的基因分型引物进行多重等位基因特异性引物延伸。然后在Tag-It阵列上对产品进行分类，并使用Luminex xMAP(TM)系统进行检测。结果：经实验验证，最高非特异性信号占特异性信号的3.7%。结论：基于Tag-it微球的通用芯片平台是一种高精度、多路、高通量的SNP检测平台。(C)2004年美国临床化学协会。
Background: We have developed a novel, microsphere-based universal array platform referred to as the Tag-It(TM) platform. This platform is suitable for high-throughput clinical genotyping applications and was used for multiplex analysis of a panel of thrombophilia-associated single-nucleotide polymorphisms (SNPs).Methods: Genomic DNA from 132 patients was amplified by multiplex PCR using 6 primer sets, followed by multiplex allele-specific primer extension using 12 universally tagged genotyping primers. The products were then sorted on the Tag-It array and detected by use of the Luminex xMAP(TM) system. Genotypes were also determined by sequencing.Results: Empirical validation of the universal array showed that the highest nonspecific signal was 3.7% of the specific signal. Patient genotypes showed 100% concordance with direct DNA sequencing data for 736 SNP determinations.Conclusions: The Tag-It microsphere-based universal array platform is a highly accurate, multiplexed, high-throughput SNP-detection platform. (C) 2004 American Association for Clinical Chemistry.