Second generation analysis of antinuclear antibody (ANA) by combination of screening and confirmatory testing

Scholz, Juliane; Großmann, Knut; Knütter, Ilka; Hiemann, Rico; Sowa, Mandy; Röber, Nadja; Rödiger, Stefan; Schierack, Peter; Reinhold, Dirk; Bogdanos, Dimitrios P.; Meroni, Pier Luigi; Radice, Antonella; Conrad, Karsten; Roggenbuck, Dirk

doi:10.1515/cclm-2015-0083

Cited by 27 publications

(22 citation statements)

References 45 publications

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“…However, IFAs have some analytical disadvantages, including subjective interpretation and limited automation. To overcome these issues, enzyme immunoassays (EIAs) were developed (8)(9)(10). Although many variations exist, most EIAs for ANA testing use an HEp-2 nuclear extract, supplemented with certain purified antigens, in place of the intact cell.…”

Section: Impact Statementmentioning

confidence: 99%

Utility of Antinuclear Antibody Screening by Various Methods in a Clinical Laboratory Patient Cohort

Duan

Peters

Ettore

et al. 2016

The Journal of Applied Laboratory Medicine

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Background Antinuclear antibody (ANA)5 testing is routinely performed during evaluation of patients with a suspected connective tissue disease (CTD), yet the question of which method is most appropriate remains controversial. The purpose of this study was to evaluate the clinical utility of ANA testing by an enzyme immunoassay (EIA), an immunofluorescence assay (IFA), and a multiplex immunoassay (MIA) in a routine laboratory population. Methods Samples (n = 1000) were collected from specimens submitted for ANA testing by EIA (Bio-Rad). All samples were subsequently analyzed by IFA (Zeus) and MIA (Bio-Rad). The sample cohort was weighted to represent the routine testing population. Diagnostic information was obtained by chart review. Results For the diagnosis of a CTD, ROC curve analysis demonstrated no significant differences between IFA (area under the curve 0.81) and EIA (0.84) (P = 0.25), with overlay of a single point for the MIA. When normalized to a specificity of approximately 90%, the sensitivities of the MIA, EIA, and IFA were 67%, 67%, and 56%, respectively. By varying the clinical cutoff, the IFA could achieve the highest sensitivity of 94%; however, the corresponding specificity was only 43%. In contrast, a strongly positive EIA had a specificity of 97%, although, at this cutoff, the sensitivity was only 40%. Conclusions Although the overall diagnostic performance of the IFA, EIA, and MIA were not statistically different, the clinical sensitivity and specificity varied dramatically based on the positive/negative cutoff. Knowledge about the performance characteristics of each method will significantly aid in the interpretation of ANA testing.

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Section: Impact Statementmentioning

confidence: 99%

Utility of Antinuclear Antibody Screening by Various Methods in a Clinical Laboratory Patient Cohort

Duan

Peters

Ettore

et al. 2016

The Journal of Applied Laboratory Medicine

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“…Antinuclear antibodies (ANA) are a heterogeneous group of autoantibodies [1] that can be detected in the sera of subjects with systemic autoimmune rheumatic diseases (SARD), such as systemic lupus erythematosus (SLE), Sjögren's syndrome (SSj), systemic sclerosis (SS), rheumatoid arthritis (RA), idiopathic inflammatory myopathies (IIM) and systemic vasculitides (SV) [2][3][4]. Autoantibodies are very helpful in the diagnosis, prognosis assessment and monitoring of the clinical evolution of patients with SARD [2,[4][5][6].…”

Section: Introductionmentioning

confidence: 99%

Predictive autoimmunity using autoantibodies: screening for anti-nuclear antibodies

Pérez

Gilburd

Cabrera-Marante³

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Background: Early detection of antinuclear antibodies (ANA) in asymptomatic subjects is useful to predict autoimmune diseases years before diagnosis. ANA have been determined by indirect immunofluorescence (IIF) using human epithelial type 2 (HEp-2) cells, which is considered the gold standard technique. Multiplex technology (BioPlex ANA Screen) has been introduced for ANA evaluation in recent years. Nevertheless, concordance between BioPlex and IIF is low and there is no harmonization between both methods for detection of autoantibodies. This study has aimed to clarify the clinical significance of autoantibodies detected by BioPlex ANA Screen in subjects with undiagnosed clinical suspicion of autoimmune disease and to determine the predictive value of autoantibodies detected by BioPlex ANA Screen. Methods: A 3-year follow-up study was performed of 411 subjects without a clear diagnosis of autoimmune diseases in whom autoantibodies were detected by BioPlex ANA Screen that were negative by IIF on HEp-2 cells.

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“…The beads coated with different antigens can be identified by the size as well as location in one out of four compartments surrounding the central part of the well containing the cellular substrate. 8,9 The Biochip method (Euroimmun, Lübeck, Germany) also enables the combination of multiple substrates in a single incubation field. These substrates may consist of any combination of cells, tissues, antigen dots, or transfected cells.…”

Section: Distinct Methods Of Multiparametric Immunoassaysmentioning

confidence: 99%

“…More recently, pattern recognition devices have become available that enable automated reading of these multiparametric IIF assays. 8,9,11 If fully automated, medium-to-high volume testing can be handled. In case of the CytoBead method, the ANCA specificity assays even generate quantitative results by utilizing a standard curve based on reference sera of the International Center for Disease Control and Prevention (CDC).…”

Section: Distinct Methods Of Multiparametric Immunoassaysmentioning

confidence: 99%

“…23 The multiparametric CytoBead assay circumvents this short coming by combining the cellular-and antigen-specific substrates. 8 In general, multiparametric assays are used as a second, or even third step in the testing algorithm for SARD. Following a positive ANA IIF result, multiparametric assays enable direct identification of possible antigen-specific antibodies.…”

Section: Multiparametric Autoantibody Diagnostics In Systemic Autoimmmentioning

confidence: 99%

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Multiparametric autoimmune diagnostics: recent advances

Damoiseaux¹

2016

PLMI

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Autoimmune diagnostics in a routine clinical laboratory is constantly challenged by the discovery of new autoantibodies and technical innovations in the immunoassays applied. These challenges are, in particular, combined in the multiparametric immunoassays. Appropriate positioning of multiparametric immunoassays within the laboratory requires integrated knowledge of the clinical performance of the test system for each individual antigen, the conditions prescribed in disease criteria and/or guidelines, and the demands of the clinicians. This review provides a summary of the multiparametric immunoassays available, as well as the applications and restrictions in routine clinical practice.

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Second generation analysis of antinuclear antibody (ANA) by combination of screening and confirmatory testing

Cited by 27 publications

References 45 publications

Utility of Antinuclear Antibody Screening by Various Methods in a Clinical Laboratory Patient Cohort

Utility of Antinuclear Antibody Screening by Various Methods in a Clinical Laboratory Patient Cohort

Predictive autoimmunity using autoantibodies: screening for anti-nuclear antibodies

Multiparametric autoimmune diagnostics: recent advances

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