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首页> 外文期刊>The journal of applied laboratory medicine. >Comparison of Next-Generation SequencingBased Human Leukocyte Antigen Typing with Clinical Flow Cytometry and Allele-Specific PCR Melting Assays for HLA-B27 Genotyping
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Comparison of Next-Generation SequencingBased Human Leukocyte Antigen Typing with Clinical Flow Cytometry and Allele-Specific PCR Melting Assays for HLA-B27 Genotyping

机译：下一代SequencingBased的比较人类白细胞抗原类型与临床流血细胞计数和Allele-Specific PCR检测融化对于HLA-B27基因分型

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Background: Due to the strong association between ankylosing spondylitis and Human Leukocyte Antigen (HLA)-B27, accurate identification of HLA-B27 is important in the diagnosis of patients with suspected spondyloar-thritides. For this study, we compared a high-resolution HLA-B typing method to the clinical flow cytometry and allele-specific PCR melting assays to determine clinical benefits of high-resolution testing. Methods: Residual clinical samples submitted for HLA-B27 testing by flow cytometry were tested by single-locus HLA-B genotyping using next-generation sequencing (NGS), and PCR with melting curve analysis, currently used as a reflex test for indeterminate flow cytometry results. Results: Fifty out of the 51 samples (98%) positive by flow cytometry confirmed as HLA-B27 positive by PCR melting assay and by NGS. The sample that did not confirm was genotyped as HLA-B*07:02. All the samples negative by flow cytometry were confirmed as HLA-B27 negative by both PCR melting assay and NGS. For the group that was indeterminate by flow cytometry, 84.5% (n = 49) typed as positive for HLA-B27, while 15.5% (n = 9) were negative for HLA-B27 but positive for HLA-B*07:02. NGS was the only method able to distinguish between pathogenic and nonpathogenic HLA-B27 variants, in contrast to the flow cytometry or the PCR melting assays. Conclusions: Single-locus NGS is superior to flow cytometry and PCR melting assay for the unambiguous identification of HLA-B27 variants, and uniquely able to distinguish between pathogenic and nonpathogenic B27 alleles. Due to its high accuracy, it may be a feasible superior alternative to flow cytometry and traditional molecular methods for clinical HIA-B27 testing
机译：背景:由于强烈的联系强直性脊柱炎和人类白细胞抗原(HLA) -B27、准确的识别HLA-B27病人的诊断是很重要的疑似spondyloar-thritides。研究中,我们比较高分辨率HLA-B打字方法临床流式细胞术和allele-specific PCR分析来确定融化临床高分辨率测试的好处。方法:残余临床样本提交HLA-B27测试通过流式细胞术进行了测试基因分型结果single-locus HLA-B使用下一代测序(门店),和PCR融化曲线分析,目前用作为不确定的流式细胞仪反射测试结果。(98%)正通过流式细胞术确认HLA-B27积极通过PCR测定融化,挥动。样品没有确认这些HLA-B * 07:02。血细胞计数被确认为HLA-B27负面两个PCR试验和门店融化。不定的流式细胞术,84.5%(n = 49)类型作为HLA-B27积极,15.5% (n = 9) - HLA-B27但HLA-B * 07:02阳性。能够区分致病性和不致病的HLA-B27变异,形成鲜明对比流式细胞术或PCR检测融化。结论:Single-locus挥动优于流血细胞计数和PCR鉴定融化明确的识别HLA-B27变异,和唯一能够区分致病性和非B27等位基因。其精度高,它可能是一个可行的优越流式细胞术和传统的替代品分子临床HIA-B27测试的方法

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