Human leukocyte antigen-B27 typing by flow cytometry: Comparison of three CE-IVD methods

IF 2.3 3区医学 Q3 MEDICAL LABORATORY TECHNOLOGY

Cytometry Part B: Clinical Cytometry Pub Date : 2022-11-30 DOI:10.1002/cyto.b.22102

Louis Waeckel, Anne Kennel, Anne-Emmanuelle Berger, Claude Lambert

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引用次数: 1

Abstract

Background

The human leukocyte antigen B27 (HLA-B27), associated with chronic inflammatory diseases is thus widely performed for diagnostic purposes. Genotyping by molecular biology is the current gold standard for HLA-B27 typing, but cheaper and faster flow cytometry methods have been developed.

Methods

In this report, we compare analytical performances of three CE-IVD flow cytometry kits: IOTest™ and DuraClone B27™ from Beckman Coulter and BD HLA-B27™ from BD Biosciences on a Navios™ cytometer as compared to molecular biology.

Results

Routine analyses could conclude for HLA-B27 in197 patients (23.2%) and was not conclusive for 66 patients (7%, 8%). The experience revealed the needs to complete IOTest™ with a lineage marker (like CD3-APC) and a standardization procedure in detection of fluorescence. Comparative analysis considering 23/44 non-conclusive samples as negative, pointed out a 100% sensitivity and specificity of IOTest™ in determining genetically proved HLA-B27. Specificity of the BD HLA-B27^TM kit was 94% (two false positives) sticking to the manufacturer instructions but raised to 100% and 94% sensitivity with a cutoff at 8.5 (225 on FACSdiva's scale).

Discussion

The DuraClone B27™ specificity was poor using the manufacturer cutoff but raised to 100% with a 8.0 cutoff instead of 15.

Conclusion

The three flow cytometry kits displayed satisfying performances but need adjustments. The DuraClone B27™ kit seems to be the best while the IOTest™ kit is not conclusive in 8% of cases. In most cases the use of molecular biology can be spared, but genotyping remains essential for patients whose HLA-B27 status cannot be determined with confidence by flow cytometry.

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通过流式细胞术进行人类白细胞抗原-B27 分型：三种 CE-IVD 方法的比较

背景与慢性炎症疾病相关的人类白细胞抗原 B27（HLA-B27）因此被广泛用于诊断目的。分子生物学基因分型是目前 HLA-B27 分型的黄金标准，但目前已开发出更便宜、更快速的流式细胞术方法。方法在本报告中，我们比较了三种 CE-IVD 流式细胞仪试剂盒的分析性能：贝克曼库尔特公司的 IOTest™ 和 DuraClone B27™ 以及 BD 生物科学公司的 BD HLA-B27™ 在 Navios™ 细胞仪上的分析性能与分子生物学方法进行了比较。结果常规分析可确定 197 名患者（23.2%）的 HLA-B27，66 名患者（7%，8%）的 HLA-B27不确定。经验表明，需要用系标记（如 CD3-APC）和标准化荧光检测程序来完成 IOTest™。比较分析将 23/44 份无结论样本视为阴性样本，结果表明 IOTest™ 在确定经基因证实的 HLA-B27 时具有 100% 的灵敏度和特异性。根据制造商的说明，BD HLA-B27TM 试剂盒的特异性为 94%（两个假阳性），但在临界值为 8.5（FACSdiva 标准为 225）时，灵敏度提高到 100%和 94%。讨论 DuraClone B27™ 的特异性较差，使用的是制造商的截止值，但在截止值为 8.0 而不是 15 时，特异性提高到了 100%。结论这三种流式细胞仪试剂盒的性能令人满意，但需要调整。DuraClone B27™ 试剂盒似乎是最好的，而 IOTest™ 试剂盒在 8%的情况下不能得出结论。在大多数情况下，可以不使用分子生物学方法，但对于无法通过流式细胞术确定 HLA-B27 状态的患者，基因分型仍是必不可少的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cytometry Part B: Clinical Cytometry 医学-病理学

CiteScore

6.80

自引率

32.40%

发文量

审稿时长

>12 weeks

期刊介绍： Cytometry Part B: Clinical Cytometry features original research reports, in-depth reviews and special issues that directly relate to and palpably impact clinical flow, mass and image-based cytometry. These may include clinical and translational investigations important in the diagnostic, prognostic and therapeutic management of patients. Thus, we welcome research papers from various disciplines related [but not limited to] hematopathologists, hematologists, immunologists and cell biologists with clinically relevant and innovative studies investigating individual-cell analytics and/or separations. In addition to the types of papers indicated above, we also welcome Letters to the Editor, describing case reports or important medical or technical topics relevant to our readership without the length and depth of a full original report.