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. 2016 Dec 20:9:1-8.
doi: 10.2147/CLEP.S121632. eCollection 2017.

Antinuclear antibody prevalence in a general pediatric cohort from Mexico City: discordance between immunofluorescence and multiplex assays

Emily C Somers  1 , Seetha U Monrad  2 , Jeffrey S Warren  3 , Maritsa Solano  4 , Lourdes Schnaas  5 , Mauricio Hernandez-Avila  4 , Martha Maria Tellez-Rojo  4 , Howard Hu  6
Affiliations

Antinuclear antibody prevalence in a general pediatric cohort from Mexico City: discordance between immunofluorescence and multiplex assays

Emily C Somers et al. Clin Epidemiol. .

Abstract

Objective: To characterize antinuclear antibody (ANA) prevalence according to distinct assay methodologies in a pediatric cohort from Mexico City, and to further examine associations with age and sex.

Methods: Serum ANA were measured by indirect immunofluorescence assay (IFA) and multiplex immunoassay in 114 children aged 9-17 years. IFA was considered positive at a cutoff titer of ≥1:80. Agreement between assay methods was assessed by kappa statistic. Sensitivity, specificity, and 95% confidence intervals (CIs) of the multiplex were computed with IFA as the reference standard.

Results: Of the 114 children (mean age 14.7 [standard deviation 2.1] years; 54 [47%] female), 18 of 114 (15.8%) were ANA positive by IFA, and 11 of 114 (9.6%) by 11-antigen multiplex assay. ANA prevalence was higher in females compared with males by both of the methods (ratios 1.6-1.9 to 1). Agreement between tests was classified as slight by kappa (κ=0.177 [95% CI -0.051, 0.406]). The multiplex immunoassay had sensitivity of 22.2% (95% CI 6.4, 47.6) and specificity of 92.7% (95% CI 85.6, 97.0), and failed to capture 3 of 4 (75%) of the high-titer (≥1:1280) IFA-positives.

Conclusion: Up to 15% of children in this general population cohort were ANA positive, with a higher rate of positivity among females according to both assay methods. Substantial discordance in ANA results was found between IFA and multiplex methods, even for high-titer IFA positives. These findings underscore the need to sufficiently account for assay characteristics when interpreting ANA test results, and support IFA as the more appropriate assay for studies of subclinical autoimmunity.

Keywords: autoantibodies; autoreactivity; biomarker; epidemiology; immune dysfunction; pediatric; preclinical; subclinical autoimmunity.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
Representative photomicrographs of antinuclear antibody (ANA) patterns at 400× magnification. (A) negative ANA; (B) positive/homogeneous pattern; (C) positive/speckled pattern; (D) positive/nucleolar pattern.
Figure 2
Figure 2
(A) ANA titers and patterns, by indirect IFA. (B) Autoantibody specificities by multiplex-11 assay. Four specificities did not yield any positives (ribosomal P, SS-A, centromere B, and Jo-1). Abbreviations: ANA, antinuclear antibody; dsDNA, double-stranded DNA; IFA, immunofluorescence assay.
Figure 3
Figure 3
Age distributions according to ANA screening status by indirect IFA and multiplex-11 assays. Abbreviations: ANA, antinuclear antibody; IFA, immunofluorescence assay.
Figure 4
Figure 4
Proportional Venn diagram displaying overlap in positivity for ANA assay methods (n=114). Corresponding sensitivity, specificity, and kappa estimates are presented, for multiplex compared to IFA (≥1:80) as the “gold standard”. Multiplex-10 excludes the dsDNA antigen from the full multiplex-11 panel. Abbreviations: ANA, antinuclear antibody; dsDNA, double-stranded DNA; IFA, immunofluorescence assay.
See this image and copyright information in PMC

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