Clinical autoantibody detection by microarray

Dillaerts, Doreen; Baere, Heidi De; Bossuyt, Xavier

doi:10.1515/cclm-2016-0533

Cited by 7 publications

(4 citation statements)

References 11 publications

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“…Recently, numerous methods have been implemented in ANA screening tests, including enzyme-linked immunosorbent immunoassay (ELISA), automated chemiluminescent immunoassay [3], microarray systems [4] and indirect immunofluorescence (IIF) assays. Although some high-throughput methods have the advantage of rapid detection, IIF using HEp-2 (human epidermoid laryngeal carcinoma) cells remains the reference technique for ANA screening, which is the initial step in diagnosing systemic autoantibodies in conditions such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjögren's syndrome (SjS) and undifferentiated connective tissue disease (UCTD) [5].…”

Section: Introductionmentioning

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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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: Introductionmentioning

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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“…In 2016, Dillaerts et al performed the AMiDot on 184 samples from blood donors and on 280 randomly selected clinical samples containing antibodies to ENAs. The κ agreement between AMiDot and FEIA was ≥0.44 (≥0.70 after exclusion of anti-SmD) [ 28 ]. The low agreement of anti-Sm was related to the fact that AMiDot uses native Sm as the antigen, but FEIA EliA uses the synthetic antigen SmD.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a New Multiparametric Microdot Array-Based Immunoassay Panel for Systemic Autoimmune Disease Diagnosis

Infantino,

Pavia,

Grossi

et al. 2024

JPM

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Background: The early reliable detection and quantification of autoantibodies play an important role in autoimmune disease diagnosis and in disease-course monitoring. New technologies, such as the multiplexed determination of autoantibodies, have recently been introduced and are being adopted more frequently. The aim of this study was to evaluate the ability of a new microdot array-based multiparametric assay (ZENIT AMiDot CTD panel, A. Menarini Diagnostics, Firenze, Italy) to correctly classify patients with autoimmune rheumatic diseases (ARDs) and compare it to a fluorescence enzyme immunoassay (FEIA) for the detection of anti-ENAs. Methods: The study included 69 consecutive samples from patients with ARDs that were analyzed using two different methods (FEIA and AMiDot) to detect anti-CENP B and six anti-ENA antibodies: anti-Scl-70, anti-SSB/La, anti-Jo-1, anti-U1-RNP, anti-Ro52, and anti-Ro60. The control group sera came from sixty-eight blood donors. Tests were run on the automated slide processor ZENIT FLOW, and then the slides were imaged and analyzed using ZENIT fast. Results: Since the samples were selected for at least one antibody positivity with an ARD diagnosis, we did not calculate clinical sensitivity but only specificity, which was 98.53%, ranging from 90% for anti-SSB/La antibodies to 100% for anti-CENP B ones. Mean agreement among the methods assessed by Cohen’s kappa was 0.816 ± 0.240. Conclusions: The assay demonstrated good clinical performance and may be considered a valuable aid in detecting ARD patients, offering an alternative to methods such as FEIA which are largely in use today.

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“…Therefore, novel automated platforms have been introduced, including chemiluminescent immunoassay (CLIA), multiplex bead-based flow fluorescent immunoassay (MBFFI), magnetic bar code immunofluorescence assay (MBC-IF), and microarray systems. [10][11][12][13][14] However, inconsistencies among methods are burdensome for those who perform and interpret the tests. 3 Inappropriate interpretation can lead to misdiagnosis, unbefitting therapies, and unnecessary costs.…”

Section: Introductionmentioning

confidence: 99%

“…Meanwhile, the demand for ANA testing has increased remarkably in recent years, in turn increasing the need for high throughput in clinical laboratories. Therefore, novel automated platforms have been introduced, including chemiluminescent immunoassay (CLIA), multiplex bead‐based flow fluorescent immunoassay (MBFFI), magnetic bar code immunofluorescence assay (MBC‐IF), and microarray systems 10–14 …”

Section: Introductionmentioning

confidence: 99%

Analytical and clinical performance of different platforms simultaneously detecting 15 antinuclear antibodies

Qin

Fan

Wang

et al. 2022

Clinical Laboratory Analysis

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Background: Antinuclear antibodies (ANAs) are invaluable biomarkers for the diagnosis of autoimmune diseases (AIDs). This study aims to compare the performances of line immunoassay (LIA), multiplex bead-based flow fluorescent immunoassay (MBFFI), and magnetic bar code immunofluorescence assay (MBC-IF) to detect ANA-Profile-15S. Methods: In total, 184 samples from AID patients and 50 healthy controls (HCs) were collected. Fifteen ANAs (anti-dsDNA, nucleosome, histone, Sm, PCNA, ribosomal-P, SS-A/Ro52, SS-A/Ro60, SS-B/La, centromere B [CENP-B], Scl-70, U1-snRNP, AMA-M2, Jo-1, and Pm/Scl) were subjected to parallel detection by the LIA, MBFFI, and MBC-IF. The consistency between assays was analyzed. The discrepant results were further examined by chemiluminescent immunoassay (CLIA). Results: Anti-SS-A/Ro52 and SS-A/Ro60 autoantibodies were the most common autoantibodies in ANA positive-profiles, and were detected with equal efficiency by the LIA, MBFFI, and MBC-IF (p = 0.101 and p = 0.732, respectively). The three assays showed excellent agreement (consistency range: 66.5%-97.5%), and total consistency was 85.8%. The MBFFI and MBC-IF assays were in good agreement in terms of ANA-Profile-15S determination; the kappa coefficient ranged from 0.59 to 0.95, except for the PCNA and PM-Scl. Of the 262 re-assessed divergent results, 124 (47.33%) were positive on CLIA; the various autoantibodies exhibited variable patterns. More importantly, the ANA-Profile-15S results of the MBFFI and MBC-IF accurately identified patients with AID; the area under the curves ranged from 0.642 to 0.919. Conclusions: The novel MBFFI and MBC-IF assay performed well in detecting ANA-Profile-15S. The application of MBFFI and MBC-IF play important roles in laboratory diagnosis of AIDs.

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Clinical autoantibody detection by microarray

Abstract: AMiDot offers an alternative to line immunodot assay. Individual antibody assays might suffer from low sensitivity.

Cited by 7 publications

References 11 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

Evaluation of a New Multiparametric Microdot Array-Based Immunoassay Panel for Systemic Autoimmune Disease Diagnosis

Analytical and clinical performance of different platforms simultaneously detecting 15 antinuclear antibodies

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