HLA-DQA1 * 05变异体（rs2097432）与炎症性肠病患者肿瘤坏死因子拮抗剂免疫原性增加和随后的治疗抵抗性相关。鉴定这些患者将优化个性化治疗选择。从未知rs2097432基因型的未选患者人群中提取了80个去标识化样本的基因组DNA。使用参考实验室进行拆分样品分析，自定义设计TaqMan定量聚合酶链反应（qPCR）特异探针（Thermo Fisher Scientific）。使用人工合成的基因组区块片段作为对照。所有qPCR反应都在Applied Biosystems 7500系统下的快速循环条件下使用TaqMan GTXpress Master Mix（Thermo Fisher Scientific）进行。在80个样本中，50%为野生型参考基因型，22.5%为杂合子，27.5%为同型变异呼叫，与人群数据类似。两个独立实验室之间的拆分分析样品完全一致。在复制处理的基因组区块对照中测试的检测限制在10ng到1.25ng的样品输入中得到复现，分别在两个独立运行中进行了测试。此外，以将前野生型参考和同型变异DNA掺入基因组区块控制中来评估进一步的分析特异性，可以产生适当的杂合基因型。我们在这里介绍了一个实验室开发的快速HLA-DQA1 * 05（rs2097432）药物基因组学检测的验证，其针对基因组范围关联研究识别的热点。此处使用的定向基因分型将允许迅速的个性化治疗选择。©2023作者。由牛津大学出版社代表美国临床病理学会出版。版权所有。有关权利，请发送电子邮件至journals.permissions@oup.com。

The HLA-DQA1*05 variant (rs2097432) is associated with increased risk of immunogenicity to tumor necrosis factor antagonists, with subsequent resistance to therapy in patients with inflammatory bowel disease. Identification of these patients would optimize personalized therapeutic selection.Genomic DNA was extracted from 80 deidentified samples in an unselected patient population with an unknown rs2097432 genotype. Split sample analysis was performed using a reference laboratory. Primer probes for a TaqMan quantitative polymerase chain reaction (qPCR) assay (Thermo Fisher Scientific) were custom designed. Synthesized genomic-block fragments were used as controls. All qPCR reactions were performed using a TaqMan GTXpress Master Mix (Thermo Fisher Scientific) on the Applied Biosystems 7500 system under fast cycling conditions.Of 80 samples, 50% were wild-type reference genotypes, 22.5% were heterozygous, and 27.5% were homozygous variant calls, comparable to population data. Split analysis samples between 2 independent laboratories were 100% concordant. The detection limit tested across genomic-block controls processed in duplicate was reproducible on sample input from 10 ng titrated down to 1.25 ng across 2 independent runs. Further, analytical specificity assessed with previous wild-type reference and homozygous variant DNA spiked into genomic-block controls produced appropriate heterozygous genotypes.Here we present validation of a lab-developed test for a rapid HLA-DQA1*05 (rs2097432) pharmacogenomics assay targeting a hotspot identified by genome-wide association studies. Targeted genotyping employed here will allow for expeditious personalized therapeutic selection.© The Author(s) 2023. Published by Oxford University Press on behalf of American Society for Clinical Pathology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.