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

Abstract

BACKGROUND:

For the serological diagnosis of systemic autoimmune rheumatic diseases, a two-tier approach starting with sensitive antinuclear antibody (ANA) detection by indirect immunofluorescence (IIF) on HEp-2 cells followed by characterization of positive findings with different immunoassays is recommended. To overcome drawbacks of this approach, we developed a novel technique allowing the combination of screening and simultaneous confirmatory testing. For the first time, this creates the basis for second generation ANA testing.

METHODS:

ANA and autoantibodies (autoAbs) to double-stranded DNA (dsDNA), CENP-B, SS-A/Ro52, SS-A/Ro60, SS-B/La, RNP-Sm, Sm, and Scl-70 were determined by IIF and enzyme-linked immunosorbent assay (ELISA), respectively, and compared to simultaneous analysis thereof by second generation ANA analysis in patients with systemic lupus erythematosus (n=174), systemic sclerosis (n=103), Sjögren's syndrome (n=46), rheumatoid arthritis (n=36), mixed and undetermined connective tissue diseases (n=13), myositis (n=21), infectious disease (n=21), autoimmune liver disease (n=93), inflammatory bowel disease (n=78), paraproteinemia (n=11), and blood donors (n=101).

RESULTS:

There was very good agreement of second generation ANA testing with classical one by IIF and ELISA regarding testing for ANA and autoAbs to dsDNA, CENP-B, SS-B, RNP-Sm, Scl-70, SS-A/Ro52, and SS-A/Ro60 (Cohen's κ>0.8). The agreement for anti-Sm autoAb was good (κ=0.77). The differences of both approaches were not significant for autoAbs to SS-B/La, RNP-Sm, Scl-70, SS-A/Ro60, and SS-A/Ro52 (McNemar's test, p>0.05, respectively).

CONCLUSIONS:

Second generation ANA testing can replace the two-tier analysis by combining IIF screening with multiplex confirmative testing. This addresses shortcomings of classical ANA analysis like false-negative ANA findings and lack of laboratory efficiency and standardization.

• Authors: Juliane Scholz, Kai Grossmann, Ilka Knütter, Rico Hiemann, Mandy Sowa, Nadja Röber, Stefan Rödiger, Peter Schierack, Dirk Reinhold, Dimitrios P Bogdanos, Pier Luigi Meroni, Antonella Radice, Karsten Conrad, Dirk Roggenbuck
• Journal: Clinical chemistry and laboratory medicine (Clin Chem Lab Med) Vol. 53 Issue 12 Pg. 1991-2002 (Nov 2015) ISSN: 1437-4331 [Electronic] Germany
• PMID: 26020561 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Antinuclear

Topics

• Antibodies, Antinuclear (blood)
• Autoimmune Diseases (blood, diagnosis, immunology)
• Fluorescent Antibody Technique, Indirect
• Hep G2 Cells
• Humans
• Immunoassay
• Rheumatic Diseases (blood, diagnosis, immunology)