HLA associations have been linked to many diseases and are important for risk assessment of drug hypersensitivity reactions. The increasing number of HLA alleles discovered generated a list of ambiguities that cannot be resolved with the current clinical assays, which commonly include sequence-specific oligonucleotide probe (SSOP) genotyping, and real-time PCR with melting curve analysis. HLA typing by next-generation sequencing (NGS) has recently been adopted by clinical laboratories for transplantation testing, as it provides unambiguous and cost-effective HLA typing. The goal of this study was to evaluate the feasibility of using NGS-based HLA-B and DQ genotyping for clinical HLA disease association testing, and provide direct comparison with the currently used clinical tests, including SSOP genotyping, and real-time PCR with melting curve analysis. While the real-time PCR method is easy and inexpensive to perform, ambiguities are rapidly increasing as more and more HLA alleles are discovered. SSOP genotyping identifies the alleles present but limitations include ambiguities and underreporting less common alleles. Our data show that HLA typing by NGS is superior to the existing clinical methods for identifying HLA alleles associated with disease or drug hypersensitivity, and offers a viable approach for high volume clinical diagnostic laboratories.

HLA关联已与许多疾病相关联，对于药物超敏反应的风险评估非常重要。发现的HLA等位基因数量不断增加，产生了一系列歧义，这些歧义无法用当前的临床检测方法解决，这些检测方法通常包括序列特异性寡核苷酸探针（SSOP）基因分型和带有熔解曲线分析的实时PCR。下一代测序（NGS）进行的HLA分型最近已被临床实验室用于移植测试，因为它提供了明确且具有成本效益的HLA分型。这项研究的目的是评估将基于NGS的HLA-B和DQ基因分型用于临床HLA疾病关联测试的可行性，并提供与当前使用的临床测试（包括SSOP基因分型，实时PCR和熔解曲线分析。尽管实时PCR方法简便易行且执行成本低廉，但随着发现越来越多的HLA等位基因，歧义迅速增加。SSOP基因分型可识别存在的等位基因，但局限性包括歧义性和少报少见的常见等位基因。我们的数据表明，通过NGS进行HLA分型优于用于鉴定与疾病或药物超敏性相关的HLA等位基因的现有临床方法，并且为大量临床诊断实验室提供了可行的方法。