Abstract:
The international high-resolution external proficiency testing (EPT) started in 2004 with high-resolution typing of human leucocyte antigen (HLA) class I (HLA-A,B,C) and HLA class II (HLA-DRB1, DRB345, DQB1, and DPB1) alleles, since possibilities for such an EPT within Europe were limited and all existing EPTs at that time made use of the comparison of HLA typing results without a reference. This EPT was set up as a collaboration between the HLA laboratory of Leiden, providing DNA samples to the participants, and the laboratory of Maastricht, performing the high-resolution typing as the reference result and evaluating the results of all participants according to the prevailing European Federation for Immunogenetics (EFI) standards. Once a year, 12 samples were sent to the participating laboratories, and evaluation and certificates were provided at the end of that same year. During the years, the EPT was extended to low-resolution HLA class I and II typing, high-resolution typing including DQA1 and DPA1, and allelic resolution typing for HLA class I, the latter one being unique in this field. Evaluation of the high-resolution typing results of the last 19 years showed a clear increase in the number of loci tested by the participating laboratories and a clear change of method from Sanger sequencing with additional other techniques (SSO/SSP) to the nowadays widely used next-generation sequencing method. By strictly using the EFI rules for high-resolution HLA typing, the participants were made aware of the ambiguities within exons 2 and 3 for class I and exon 2 for class II and the presence of null alleles even in a two-field HLA typing. There was an impressive learning curve, resulting in >98% correctly typed samples since 2017 and a 100% fulfillment of EFI rules for all laboratories for all loci submitted in the last 2 years. Overall, this EPT meets the need of an EPT for high-resolution typing for EFI accreditation.
摘要:
国际高分辨率外部能力测试(EPT)始于2004年,对人类白细胞抗原(HLA)I类(HLA-A,B,C)和HLAII类(HLA-DRB1,DRB345,DQB1和DPB1)等位基因,因为在欧洲进行这种EPT的可能性有限,并且当时所有现有的EPT都使用了HLA分型结果的比较,而没有参考。这个EPT是由莱顿的HLA实验室合作建立的,向参与者提供DNA样本,还有马斯特里赫特的实验室,进行高分辨率分型作为参考结果,并根据现行的欧洲免疫遗传学联合会(EFI)标准评估所有参与者的结果。一年一次,12个样本被送到参与的实验室,同年年底提供了评估和证书。多年来,EPT扩展到低分辨率HLAI类和II类分型,包括DQA1和DPA1在内的高分辨率分型,以及HLAI类的等位基因分辨率分型,后者在这个领域是独一无二的。对过去19年的高分辨率分型结果的评估表明,参与实验室测试的基因座数量明显增加,并且从Sanger测序与其他技术(SSO/SSP)到如今广泛使用的方法发生了明显变化。下一代测序方法。通过严格使用EFI规则进行高分辨率HLA分型,参与者意识到I类的外显子2和3以及II类的外显子2内的歧义,并且即使在两域HLA分型中也存在无效等位基因.有一个令人印象深刻的学习曲线,自2017年以来,超过98%的样本正确分型,并且在过去2年中提交的所有基因座的所有实验室均100%满足EFI规则。总的来说,此EPT满足EFI认证的高分辨率打字EPT的需要。