Testing for myositis specific autoantibodies: Comparison between line blot and immunoprecipitation assays in 57 myositis sera

Ilaria Cavazzana, Micaela Fredi, Angela Ceribelli, Cristina Mordenti, Fabio Ferrari, Nice Carabellese, Angela Tincani, Minoru Satoh, Franco Franceschini

Research output: Contribution to journalArticlepeer-review


Objective: To analyze the performance of a line blot assay for the identification of autoantibodies in sera of patients affected by myositis, compared with immunoprecipitation (IP) as gold standard. Methods: 66 sera of patients with myositis (23 polymyositis, 8 anti-synthetase syndromes, 29 dermatomyositis and 6 overlap syndromes) were tested by commercial LB (Euroimmun, Lubeck, Germany); 57 sera were analyzed also by IP of K562 cell extract radiolabeled with 35S-methionine. Inter-rater agreement was calculated with Cohen's k coefficient. Results: Myositis-specific antibodies (MSA) were detected in 36/57 sera (63%) by IP and in 39/66 sera (59%) by LB. The most frequent MSA found by LB were anti-Jo1 and anti-Mi2 found in 15% (10/66) of sera, followed by anti-NXP2 and anti-SRP detected in 106% (7/66) of sera. Anti-TIF1gamma and anti-MDA5 were found in 6 (9%) and 5 sera (7.6%), respectively.A good agreement between methods was found only for anti-TIF1γ, anti-MDA5 and anti-NXP-2 antibodies, while a moderate agreement was estimated for anti-Mi2 and anti-EJ. By contrast, a high discordance rate for the detection of anti-Jo1 antibodies was evident (k: 0.3).Multiple positivity for MSA were found in 11/66 (17%) by LB and 0/57 by IP (p: 0001). Comparing the clinical features of these 11 sera, we found total discrepancies between assays in 3 sera (27.3%), a relative discrepancy due to the occurrence of one discordant autoantibody (not confirmed by IP) in 5 cases (45.5%) and a total discrepancy between LB and IP results, but with a relative concordance with clinical features were found in other 3 sera (27.3%). The semiquantitative results do not support the interpretation of the data. Conclusions: The use of LB assay allowed the detection of new MSA, such as anti-MDA5, anti-MJ and anti-TIF1gamma antibodies, previously not found with routine methods. However, the high prevalence of multiple positivities and the high discondant rate of anti-Jo1 antibodies could create some misinterpretation of the results from the clinical point of view. These data should be confirmed by enlarging the number of myositis cases.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalJournal of Immunological Methods
Publication statusPublished - Jun 1 2016


  • Anti-Jo1
  • Anti-synthetase syndrome
  • Autoantibodies
  • Immunoprecipitation
  • Line blot
  • Myositis

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy


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