Anticardiolipin antibody assay: a methodological analysis for a better consensus in routine determinations - A cooperative project of the European Antiphospholipid Forum

Despite' the widely recognized practical importance of anticardiolipin (aCL ) ELISA, the reliability of this test has been recently discussed. In order to investigate this area on European scale, we sent to 30 experienced cent ers a questionnaire focusing on the diagnostic procedures applied to patien ts with antiphospholipid syndrome (APS) and on the detailed protocols used to perform aCL. Anticardiolipin ELISA was found to be the most frequently p erformed test in patients with suspected APS, but significant difference wa s shown among the various protocols. The cross-laboratory multiple examinat ion of ten serum samples evaluated independently by the 24 centers pointed out the difficulty in getting comparable results. Therefore a "consensus" p rotocol was derived from the aCL methods giving the best performance. The m aterials and reagents necessary to perform the "consensus" method, includin g, as putative standards, one IgG and one IgM monoclonal antibody (HCAL and EY2C9) were distributed to 19 Centers. The results of one IgG and one IgM aCL high positive sera measured in serial dilutions were compared. A progre ssive decrease in the variability of the values obtained for a given sample appeared evident when all the laboratories used the same standard, in thei r own in-house ELISA and even more in the "consensus" ELISA. Our data show that aCL ELISA standardization is necessary in order to obtai n comparable results in different laboratories.