Detecção de anticorpos antinucleares por imunofluorescência indireta em células HEp-2: definindo a diluição de triagem adequada para o diagnóstico das doenças reumáticas autoimunes

Detecção de anticorpos antinucleares por imunofluorescência indireta em células HEp-2: definindo a diluição de triagem adequada para o diagnóstico das doenças reumáticas autoimunes

Advances in Rheumatology, January 2014

24878786

Fabiano de Almeida Brito, Silvana Maria Elói Santos, Gilda Aparecida Ferreira, William Pedrosa, Janaina Gradisse, Lara Cristina Costa, Suzane Pretti Figueiredo Neves

To establish the abnormal title and the appropriate screening dilution for ANA (antinuclear antibodies) test by indirect immunofluorescence on HEp-2 cells (ANA HEp-2). An analysis of ANA Hep-2 in serum samples from 126 healthy individuals was performed. The samples were screened at a dilution of 1:80, and those positive were diluted to the title of 1:5120. The abnormal title of ANA was defined as that corresponding to the 95th percentile of the test in this population. The sensitivity of the different titles of antinuclear antibodies was determined in a group of 136 patients with a diagnosis of autoimmune rheumatic disease, and the specificity was determined in a group of 118 patients with other rheumatic diseases. The optimal cutoff value of the test was determined by ROC curve analysis. The frequency of ANA positivity in healthy subjects was 13.2%. There was no difference in the frequency of positive results according to gender or age. The abnormal title of ANA was defined as the dilution of 1:160. The 1:80 dilution had sensitivity of 87.7% and specificity of 67.8%, while the 1:160 dilution had sensitivity of 82% and specificity of 73.7%. By ROC curve analysis, a dilution of 1:160 corresponded to the optimal cutoff value. The abnormal title and the optimal cutoff value of ANA HEp-2 in the population was 1:160. Therefore, the dilution of 1:160 is the optimal screening dilution, with better specificity but without significantly compromising the sensitivity of the diagnostic test.