Automated indirect immunofluorescence antinuclear antibody analysis is a standardized alternative for visual microscope interpretation
Automated indirect immunofluorescence antinuclear antibody analysis is a standardized alternative for visual microscope interpretation
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DOI：
10.1515/cclm-2013-0016
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发表时间：
2013-09-01
影响因子：
6.8
通讯作者：
Devreese, Katrien
中科院分区：
文献类型：
--
作者：
Bonroy, Carolien;Verfaillie, Charlotte;Devreese, Katrien
背景：抗核抗体（ANA）筛查是全身性风湿性疾病血清学检查的基本工具。尽管出现了替代的筛查方法和标准化的困难，间接免疫荧光（IIF）仍然是ANA检测的推荐方法。本研究旨在评估自动ANA IIF分析作为传统手动方法的标准化替代方法的可靠性。方法：对304个连续的常规血清、28个显示罕见染色模式的血清库样品、219个明确定义的疾病组样品[141个系统性硬化症（SSc）、13个风湿性多肌痛、22个骨关节炎、23个关节炎、24个关节炎]进行HEp-2000细胞的ANA测试。5例ANCA相关血管炎和38例脊柱关节炎]和100例健康供体。所有样本均通过自动化IIF（Zenit G-sight）、常规目视IIF显微镜和两种ANA筛选酶免疫分析（EIA）进行分析。结果：自动化和常规ANA IIF分析在阴性/阳性解释以及强度评估方面具有可比性（>90%一致性）。相比之下，模式识别的准确性（26%）是有限的。SSc在Zenit G-sight和EIA结果区间的似然比（LR）随阳性水平的增加而增加。Zenit G-sight（97%-100%）的Ssc相关抗体亚群的灵敏度高于EIA（10%-96%）。Zenit G-sight获得的定量结果与常规终点滴度之间存在显著相关性。结论：Zenit G-sight用于ANA IIF自动分析提供了提高标准化的机会。然而，专家技术人员的补充作用仍然存在，特别是在模式识别和不确定/阴性样本分类方面。
Background: Screening for antinuclear antibodies (ANA) is a basic tool in the serological work-up of systemic rheumatic disorders. Despite the emergence of alternative screening methods and the difficulties in standardization, indirect immunofluorescence (IIF) remains the recommended method for ANA detection. This study aimed to assess the reliability of automated ANA IIF analysis as a standardized alternative for the conventional manual approach.Methods: ANA testing on HEp-2000 cells was performed on 304 consecutive routine sera, 28 serumbank samples displaying rare staining patterns, 219 samples of well-defined disease cohorts [141 systemic sclerosis (SSc), 13 polymyalgia rheumatica, 22 osteoarthritis, 5 ANCA-associated vasculitis and 38 spondyloarthritis] and 100 healthy donors. All samples were analyzed by automated IIF (Zenit G-sight), by conventional visual IIF microscopy and two ANA screening enzyme immunoassays (EIA).Results: Automated and conventional ANA IIF analysis were comparable for negative/positive interpretation as well as intensity assessment (>90% agreement). In contrast, the accuracy of pattern recognition (26%) was limited. Likelihood ratios (LR) for SSc on results intervals of both Zenit G-sight and EIA increased with increasing level of positivity. Sensitivity within the SSc-associated antibody subsets was higher for Zenit G-sight (97%-100%) than EIA (10%-96%). A significant correlation between the quantitative result obtained by Zenit G-sight and the conventional end-point titer was found.Conclusions: The use of Zenit G-sight for automated ANA IIF analysis offers opportunities to improve standardization. However, a complementary role of the expert technicians remains, especially for pattern recognition and classification of uncertain/negative samples.