Antinuclear Antibody (ANA) with HEp-2 Substrate

Antinuclear Antibody (ANA) with HEp-2 Substrate, IgG by IFA with Reflex by Pattern 3000601
Method: Semi-Quantitative Indirect Fluorescent Antibody/Qualitative Enzyme-Linked Immunosorbent Assay/Semi-Quantitative Enzyme-Linked Immunosorbent Assay/Semi-Quantitative Multiplex Bead Assay/Qualitative Immunoblot

Initial screen for autoimmune connective tissue diseases

One or more reflexive tests may be added, depending on ANA pattern detected (see ANA IFA Reflex Testing Algorithm)

Antinuclear Antibody (ANA) with HEp-2 Substrate, IgG by IFA 3000082
Method: Semi-Quantitative Indirect Fluorescent Antibody
Antinuclear Antibodies (ANA), IgG by ELISA with Reflex to ANA, HEp-2 Substrate, IgG by IFA 0050080
Method: Qualitative Enzyme-Linked Immunosorbent Assay/Semi-Quantitative Indirect Fluorescent Antibody
Antinuclear Antibodies (ANA), IgG by ELISA with Reflex to ANA HEp-2 Substrate, IgG by IFA and ENA Confirmation 0050317
Method: Qualitative Enzyme-Linked Immunosorbent Assay/Semi-Quantitative Indirect Fluorescent Antibody/Semi-Quantitative Multiplex Bead Assay

For other reflex tests, panel tests, and single tests, see Related Tests

Antinuclear antibody (ANA) testing is used in the diagnostic evaluation of various autoimmune diseases, including connective tissue diseases such as systemic lupus erythematosus (SLE), Sjögren syndrome, and systemic sclerosis (SSc). Initial testing for autoimmune connective tissue diseases (also referred to as systemic autoimmune rheumatic diseases, or SARDs) should include C-reactive protein (CRP), ANA, rheumatoid factor, and cyclic citrullinated peptide antibodies. Follow-up or confirmatory testing for positive ANA may be guided pattern(s) observed and/or patient’s clinical presentation.

Disease Overview

Diagnostic Issues

Autoimmune connective tissue diseases may present with similar features, making diagnosis difficult. Possible diagnoses may include:

ANA with reflex by immunofluorescent assay (IFA) (based on ANA patterns) may help in distinguishing between diseases.

Pathophysiology

Antigen/antibody complexes affect a variety of organs in connective tissue diseases, which frequently leads to a multisystem disease presentation. ANA antibodies are the most common antibodies and may precede the onset of connective tissue disease. While certain antibodies may show specificity for certain diseases (eg, SSA 52, SSA 60, and SSB antibodies for Sjögren syndrome), ANA antibodies are not specific for connective tissue disease and may also be associated with infectious diseases, cancers, other autoimmune disorders (eg, autoimmune liver disease), and advanced age, and may even be present in healthy patients.

Test Interpretation

Results

Dual or mixed pattern may indicate disease overlap. Visit the International Consensus on Antinuclear Antibody Patterns website  for additional information about pattern and disease associations.

Limitations

  • Dual or mixed patterns will not be reflexed; additional testing for dual or mixed patterns should be determined by the ordering physician
  • A negative ANA by IFA test does not rule out the presence of connective tissue disease

ANA IFA Reflex Testing Algorithm

References 
  1. International Consensus on Antinuclear Antibody Patterns. Nuclear Patterns. International Consensus on ANA Patterns. Gainesville, FL [Updated: 2019; Accessed: Sep 2019]
  2. Tebo AE. Recent approaches to optimize laboratory assessment of antinuclear antibodies. Clin Vaccine Immunol. 2017; 24(12): PubMed

Last Update: November 2019