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Updated: Aug 15, 2026

Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
Prevalence of positive antinuclear antibodies in healthy children
Siriwan Wananukul1, Wanlapa Voramethkul, Yadah Kaewopas
1Division of Pediatric Dermatology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. wananusw@hotmail.com
Insights
The prevalence of antinuclear antibodies (ANA) in healthy children is 15% at a 1:40 dilution or higher. This finding is crucial for accurately diagnosing connective tissue diseases in pediatric patients.
Area of Science:
- Pediatric Rheumatology
- Immunology
- Clinical Diagnostics
Background:
- Antinuclear antibodies (ANA) are key biomarkers for diagnosing pediatric connective tissue diseases.
- Understanding the prevalence of ANA in healthy children is essential for accurate diagnostic interpretation.
Purpose of the Study:
- To determine the prevalence of antinuclear antibodies (ANA) in a healthy pediatric population.
- To establish baseline ANA levels for comparison with disease states.
Main Methods:
- Indirect immunofluorescence assay using HEp-2 cells was performed on 207 serum samples from healthy children.
- Serum samples were tested for ANA at dilutions of 1:40 and higher.
- ANA results were compared with those from 52 pediatric Systemic Lupus Erythematosus (SLE) patients.
Main Results:
- Antinuclear antibodies were detected in 15% of healthy children at a dilution of 1:40 or higher.
- Specific prevalence rates in healthy children were 9% at 1:40, 3% at 1:80, and 3% at 1:160.
- ANA positivity in pediatric SLE patients was 91%, with varying titers.
Conclusions:
- The prevalence of ANA in healthy children, using HEp-2 cells and a 1:40 dilution cutoff, is 15%.
- The test demonstrated high sensitivity (91%) and negative predictive value (97%) for ANA detection.
- This study provides critical data for interpreting ANA results in pediatric autoimmune disease evaluations.
Abstract:
Antinuclear antibodies (ANA) frequently arise in the sera of children with connective tissue disease and is used in the diagnosis of these diseases. Therefore it is also important to know the prevalence of ANA in normal children. The main objective of the present study was to determine the prevalence of antinuclear antibody (ANA) in healthy children. Ninety-nine serum samples from a serum bank and 108 samples from patients who had attended elective surgery and whose blood had been withdrawn for other investigations, were tested for ANA by indirect immunofluorescence method using HEp-2 cells as substrate. Sera from 52 children with SLE were also tested during the same period. It was found that antinuclear antibodies were present in 32 (15%) of the 207 sera of healthy children at a dilution of 1:40 or higher. ANA were positive in 9% at a serum dilution of 1:40, in 3% at 1:80 and in 3% at 1:160. The patterns of immunofluorescence staining were as follows: homogeneous in 46.7%, speckled in 20%, and nucleolar in 10%. In SLE patients, ANA were positive in 91%; 13% at a serum dilution of 1:40, 7% at 1:80, 20% at 1:160, 15% at 1:320, 9% at 1:640, 20% at 1:1,280 and 9% at > or = 1:2,560. It was concluded that the prevalence of positive ANA using the HEp-2 cells as substrate was 15% in healthy children at dilutions of 1:40 or higher. Using the cutoff serum dilution of 1:40, the sensitivity of this test was 91%, the specificity was 85%, the positive predictive value was 57% and the negative predictive value was 97%.
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