Interference of Antinuclear Antibody (ANA) in Indirect Immunofluorescence Assay (IIFA)-Based Perinuclear Antineutrophil Cytoplasmic Antibody (pANCA) Interpretation.

IF 1.7 Q4 IMMUNOLOGY

Autoimmune Diseases Pub Date : 2022-10-26 eCollection Date: 2022-01-01 DOI:10.1155/2022/1343805

Sangeeta Deka, Deepjyoti Kalita, Udaykumar Sasi Rekha, Putul Mahanta, Diksha Rani, Ravi Shankar, Anusha Krishna Raj, Mithilesh Kumar Jha, Gaurav Badoni, Manisha Paul, Shailesh Kumar Gupta, Shailender Negi, Anshu Singh, Kuhu Chatterjee

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Abstract

Background: Indirect immunofluorescence assay (IIFA) based on antineutrophil cytoplasmic antibody (ANCA) testing is a commonly employed test for diagnosing autoimmune vasculitis. Antinuclear antibody (ANA) can give rise to a false interpretation of perinuclear-ANCA (pANCA) in ethanol-fixed granulocyte substrates. Analytical interference could frequently occur in setups where ethanol-fixed substrates are used alone. Here, we intend to investigate this ANA interference in pANCA interpretation.

Methods: In this retrospective study, we studied anti-MPO-negative but ANA-positive and pANCA (IIFA based) samples. We also correlated immunoblot results (where data were available) and checked the association between grades of blot positivity (an indicator of the concentration of ANA) and frequency of pANCA interpretation. Data were analyzed by appropriate statistical techniques (Chi-square and kappa statistics).

Results: About 19.2% of ANA blot (ENA-blot) positive samples displayed a pANCA positive pattern in the ethanol-fixed substrate, while this positivity in ENA-blot negatives was 6.5%. In positive ANA-IIFA samples, about 14.7% yielded pANCA patterns (on ethanol fixed substrates). Out of this, nuclear homogenous pattern yielding samples gave the highest frequency pANCA, that is, in 31.5% followed by speckled (11.1%), DFS (10.3%), and centromere (6.7%).The association of the nuclear homogenous pattern was statistically significant.

Conclusions: ANA-positive results may interfere with the interpretation of pANCA as observed in ANA-IIFA and ENA-blot positive samples. ANA-IIFA patterns like nuclear homogenous may strongly associate this pANCA interpretation. This can help laboratories perform ANCA testing more effectively, ruling out ANA interference in ANCA screening.

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基于间接免疫荧光法(IIFA)的核周抗中性粒细胞胞浆抗体(pANCA)解释中抗核抗体(ANA)的干扰

背景:基于抗中性粒细胞胞浆抗体(ANCA)检测的间接免疫荧光法(IIFA)是诊断自身免疫性血管炎的常用方法。抗核抗体(ANA)可引起对乙醇固定粒细胞底物中核周anca (pANCA)的错误解释。在单独使用乙醇固定底物的装置中，分析干扰可能经常发生。在这里，我们打算研究这种ANA对pANCA解释的干扰。方法:在这项回顾性研究中，我们研究了抗mpo阴性但ana阳性和pANCA(基于IIFA)的样本。我们还将免疫印迹结果(在有数据的情况下)进行了关联，并检查了印迹阳性等级(ANA浓度的指标)与pANCA解释频率之间的关系。采用适当的统计学方法(卡方统计和kappa统计)对数据进行分析。结果:在乙醇固定底物中，约19.2%的ANA blot (ENA-blot)阳性样品显示出pANCA阳性模式，而在ENA-blot阴性样品中，该阳性模式为6.5%。在阳性的ANA-IIFA样品中，约14.7%产生pANCA图案(在乙醇固定底物上)。其中，核均质模式产生的样品的pANCA频率最高，为31.5%，其次是斑点(11.1%)，DFS(10.3%)和着丝粒(6.7%)。核同质模式的关联具有统计学意义。结论:在ANA-IIFA和ENA-blot阳性样本中观察到，ana阳性结果可能干扰对pANCA的解释。类似核同质的ANA-IIFA模式可能与这种pANCA解释密切相关。这可以帮助实验室更有效地进行ANCA检测，排除ANA对ANCA筛查的干扰。

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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