Value-added reporting of antinuclear antibody testing by automated indirect immunofluorescence analysis

Schouwers, Sofie; Bonnet, Myriam; Verschueren, Patrick; Westhovens, René; Blockmans, Daniel Engelbert; Mariën, Godelieve; Bossuyt, Xavier

doi:10.1515/cclm-2013-0610

Cited by 27 publications

(22 citation statements)

References 18 publications

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“…Our results showed a significant association between the LIU value and the end-point titer at a dilution of 1:80, which was consistent with the report of S. Schouwers et al [17,25]. However, we further proved that the custom cutoff was superior to the preset cutoff in NOVA View (local vs. preset cutoff: 57.4% vs. 43.7%, respectively, p < 0.01) in our laboratory.…”

Section: Discussionsupporting

confidence: 93%

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Automated antinuclear immunofluorescence antibody analysis is a reliable approach in routine clinical laboratories

Zheng

Zhu

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Background: Indirect immunofluorescence (IIF) assays are recommended as the gold standard method for the detection of antinuclear antibodies (ANAs). This study aimed to investigate the reliability of an automated system. Methods: We compared 3745 serum samples using NOVA View archived images with manual analysis via microscopy. A custom cutoff value was established to distinguish ANA titers and was validated in two clinical laboratories. The automatic ANA pattern recognition system was evaluated, and all ANA-positive sera were subjected to two commercial ANA IIF kits to compare the consistency of the pattern interpretation results. For inconsistent patterns, a third ANA IIF testing kit was utilized. Results: Agreement of the interpretation of the ANA IIF test using the platform of NOVA View and manual microscopy was 96.9%. The local cutoff value to discriminate ANA titers in four main ANA patterns was calculated based on 1390 serum samples. In our laboratory, the titer prediction accuracy was superior to the preset cutoff in NOVA View (p < 0.01); the performance was similar in another laboratory (p = 0.11). The automatic pattern recognition accuracies of speckled, homogeneous, centromere, nucleolar and nuclear dot patterns were 62.7%, 57.4%, 92.6%, 30.5% and 27.3%, respectively. The consistency of the pattern interpretation results between INOVA and MBL kits was 95.3%. Conclusions: It is necessary to establish a custom valueadded ANA report. However, confirmation of the digital immunofluorescence images by expert technicians was essential, and suspect results of an ANA pattern should be reconfirmed by another commercial ANA IIF kit to achieve more reliable results.

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Section: Discussionsupporting

confidence: 93%

“…There is evidence to indicate that a high ANA titer is more related to autoimmune diseases [2,24,25]. However, with respect to clinical efficacy, the endpoint titer of ANA is not commonly manipulated in clinical laboratories in routine work.…”

Section: Discussionmentioning

confidence: 99%

Automated antinuclear immunofluorescence antibody analysis is a reliable approach in routine clinical laboratories

Zheng

Zhu

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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“…For many AARD, the likelihood for disease increases with increasing antibody concentration [28–30]. This information is lacking when a single cut-off is used.…”

Section: Statistical Considerationsmentioning

confidence: 99%

“…The currently available automated ANA IIF image analyzing systems include NOVA View (INOVA Diagnostics, San Diego, US) [30], Aklides (Medipan, Berlin, Germany) [15, 16, 75], G-Sight (Menarini, Florence, Italy) [76, 77], EuroPattern (Euroimmun, Lübeck, Germany) [73], Image Navigator (ImmunoConcepts, Sacramento, US), and Helios (Aesku, Wendelsheim, Germany) (Table 7). The systems differ from each other with respect to the use of DNA-binding counterstains, such as DAPI, the cell substrate used (e.g., most systems are restricted to using the respective manufacturer's slides), the throughput, the number of patterns that can be identified, and user-friendly features of the software [76, 77].…”

Section: Screening and Profile Assays For Ana Detectionmentioning

confidence: 99%

Current Concepts and Future Directions for the Assessment of Autoantibodies to Cellular Antigens Referred to as Anti-Nuclear Antibodies

Mähler

Meroni

Bossuyt

et al. 2014

Journal of Immunology Research

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159

151

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The detection of autoantibodies that target intracellular antigens, commonly termed anti-nuclear antibodies (ANA), is a serological hallmark in the diagnosis of systemic autoimmune rheumatic diseases (SARD). Different methods are available for detection of ANA and all bearing their own advantages and limitations. Most laboratories use the indirect immunofluorescence (IIF) assay based on HEp-2 cell substrates. Due to the subjectivity of this diagnostic platform, automated digital reading systems have been developed during the last decade. In addition, solid phase immunoassays using well characterized antigens have gained widespread adoption in high throughput laboratories due to their ease of use and open automation. Despite all the advances in the field of ANA detection and its contribution to the diagnosis of SARD, significant challenges persist. This review provides a comprehensive overview of the current status on ANA testing including automated IIF reading systems and solid phase assays and suggests an approach to interpretation of results and discusses meeting the problems of assay standardization and other persistent challenges.

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“…[21][22][23] The systems differ from each other with respect to the use of DNA-binding counterstains, such as DAPI, the cell substrate used, the throughput, the number of patterns that can be identified, and user friendly Generally, these automated systems are based on a microscope fitted with an automated stage, a CCD digital camera, a LED light source, and software that controls the moving parts and directs image acquisition. All systems perform some kind of fluorescent light intensity measurement and use the results for preliminarily categorization of the samples as positive or negative and for pattern analysis.…”

Section: 20mentioning

confidence: 99%

Anti-Nuclear antibodies: Current concepts and future direction for diagnosing connective tissue disease

Gautam¹

2015

J Pathol Nep

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Identification of antinuclear antibodies has been used for the diagnosis of connective tissue diseases for more than fifty years. Indirect immunofluorescence on human epithelial (HEp-2) cells is considered the gold standard screening method for the detection of antinuclear autoantibodies. As the demand of ANA testing increased, the need for automation and standardization has also come forth. A high level of false positive and false negative cases is seen in various populations making it difficult to take clinical decisions. Newer technologies were introduced for the antibody detection to ensure high sensitivity and specificity. This article intends to provide an overview of the concepts on ANA testing, the different diagnostic methods available, the various patterns and clinical utility, the clinical guidelines to be followed, the drawbacks and what lies ahead in the future of ANA testing.

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Value-added reporting of antinuclear antibody testing by automated indirect immunofluorescence analysis

Cited by 27 publications

References 18 publications

Automated antinuclear immunofluorescence antibody analysis is a reliable approach in routine clinical laboratories

Automated antinuclear immunofluorescence antibody analysis is a reliable approach in routine clinical laboratories

Current Concepts and Future Directions for the Assessment of Autoantibodies to Cellular Antigens Referred to as Anti-Nuclear Antibodies

Anti-Nuclear antibodies: Current concepts and future direction for diagnosing connective tissue disease

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