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Jornal Brasileiro de Patologia e Medicina Laboratorial
On-line version ISSN 1678-4774
MELLO, Rômulo Carvalho Vaz de et al. Comparative analysis of four ANA HEp-2 kits for the detection of serum autoantibodies. J. Bras. Patol. Med. Lab. [online]. 2012, vol.48, n.3, pp.185-193. ISSN 1678-4774. http://dx.doi.org/10.1590/S1676-24442012000300006.
INTRODUCTION: The objective of this study was to assess the concordance among antinuclear antibody (ANA) HEp-2 kits as to the observed fluorescent patterns. The objective was also to evaluate their performance concerning the following items: fluorescence intensity due to the use of fluorescent conjugates, the number of mitosis per field on the reaction wells, and the required dilution to obtain minimum reactivity (1+) of the reagent control-serum present in the kits. MATERIAL AND METHODS: Seventy-four samples previously known results were tested using four commercial kits ANA HEp-2 kits. The statistical evaluation was carried out with STATA® 9.2 software and the concordance among the results was assessed with Kappa test. RESULTS: The concordance coefficient among the kits obtained with Kappa test was higher than 0.94. There was a great variability as to the number of mitosis per field on the reaction wells and the intensity of fluorescence conjugates. Concerning the required dilution to obtain a minimum reactivity (1+), there was a significant difference in two kits. CONCLUSION: The general concordance level obtained with Kappa test was higher than 0.94. The greatest differences were observed in samples with nuclear fine speckled (FS), nuclear dense fine speckled (DFS) and nuclear homogeneous (HOMO) patterns. As far as serum for minimum reactivity level and intensity of fluorescent conjugates are concerned, a better standardization is yet to be pursued.
Keywords : Autoantibodies; Antinuclear antibodies; ANA HEp-2; Autoimmune diseases.