Quantification of autoantibodies to annexin V in plasma by an "in house" sandwich ELISA

Journal of the Brazilian Chemical Society

Héctor F. Pelusa²  María De Los Ángeles Valdés²  Stella M. Daniele²  Adriana M. Almará²  María J. Svetaz¹  Mariela Bearzotti¹  Cecilia V. Paparella¹  Beatriz R. Bouvet¹  Sergio A. Ghersevich²  Sandra M. Arriaga² 

[1] ,Universidad Nacional de Rosario Facultad de Ciencias Bioquímicas y Farmacéuticas Área Bioquímica ClínicaRosario,Argentina

关键词: ELISA;    annexin V;    antibodies;    anti-phospholipid syndrome;    foetal losses;   

DOI  :  10.1590/S0103-50532012000300011

来源: SciELO

【 摘 要 】

An "in house" sandwich ELISA for the quantification of plasma anti-annexin V antibodies was developed and validated. The validation parameters studied were: (i) precision, expressed either as the intra- or the inter-assay coefficient of variation (CV), (ii) exactitude, expressed as the percentage deviation between the obtained value and the real value, (iii) limit of detection (LOD), evaluated from the reagents blank and (iv) robustness, obtained by deliberately introducing slight variations in different parameters. Also, a comparison between the "in house" technique and a commercial method was performed. The research revealed that both CV were < 20%, exactitude was within the 100 ± 20% range, limit of detection was below 1 U mL-1 and that slight variations in the technique did not produce any significant variations in the results. Comparison with the commercial method showed an acceptable correlation. It was concluded that the method developed here satisfactorily accomplishes the parameters of standardization and validation for an immunoassay.

【 授权许可】

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