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Updated: Aug 9, 2026

Anti-Nuclear Antibody Screening Using HEp-2 Cells
13:01

Anti-Nuclear Antibody Screening Using HEp-2 Cells

Published on: June 24, 2014

Comparison of antinuclear antibody testing methods: immunofluorescence assay versus enzyme immunoassay

R A Gniewek1, D P Stites, T M McHugh

  • 1Bio-Rad Laboratories, Hercules, California 94547, USA.

Clinical and Diagnostic Laboratory Immunology
|March 1, 1997
PubMed

Insights

Enzyme immunoassay (ANA-EIA) for anti-nuclear antibodies shows similar sensitivity but improved specificity compared to immunofluorescence assay (ANA-IFA). This suggests ANA-EIA is a more accurate diagnostic tool for connective tissue diseases.

Area of Science:

  • Immunology
  • Clinical Chemistry
  • Autoimmunity

Background:

  • Anti-nuclear antibody (ANA) testing is crucial for diagnosing autoimmune and connective tissue diseases.
  • Immunofluorescence assay (ANA-IFA) and enzyme immunoassay (ANA-EIA) are common methods for ANA detection.
  • Comparative performance data between ANA-IFA and ANA-EIA in relation to patient diagnosis is essential for clinical practice.

Purpose of the Study:

  • To compare the diagnostic performance of ANA-IFA and ANA-EIA.
  • To evaluate sensitivity, specificity, and predictive values of both assays in a patient cohort.
  • To determine the optimal assay for ANA detection in clinical settings.

Main Methods:

  • A total of 467 patient serum samples were analyzed using both ANA-IFA (Kallestad; Sanofi) and ANA-EIA (RADIAS; Bio-Rad).
  • Patient data including age, sex, diagnosis, disease status, and medications were collected via chart review.
  • Reference ranges were established using 98 healthy blood donor samples.

Main Results:

  • ANA-EIA demonstrated higher specificity (60.2%) compared to ANA-IFA (48.0%).
  • Sensitivity was comparable between ANA-EIA (90.7%) and ANA-IFA (87.2%).
  • Positive predictive value was 35.8% for ANA-EIA and 29.1% for ANA-IFA, while negative predictive values were 96.4% and 93.9%, respectively.

Conclusions:

  • ANA-EIA offers equivalent sensitivity and superior specificity to ANA-IFA for anti-nuclear antibody testing.
  • The findings support the use of ANA-EIA as a potentially more accurate and reliable method for diagnosing connective tissue diseases.

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