A comparative study of immunological methods for determination of serum antinuclear antibodies
| Autor: | Marian M. Draganov, Mariana A. Murdjeva, Lyubomir D. Paunov, Nadezhda P. Ryasheva |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
musculoskeletal diseases
Indirect immunofluorescence Anti-nuclear antibody biology business.industry Immunoblotting Significant difference Enzyme-Linked Immunosorbent Assay General Medicine Gold standard (test) Serum samples Standard technique Molecular biology Cell Line stomatognathic diseases Specific antibody immune system diseases Antibodies Antinuclear biology.protein Humans Medicine Antibody Fluorescent Antibody Technique Indirect skin and connective tissue diseases business |
| Zdroj: | Folia Medica. 53:21-27 |
| ISSN: | 1314-2143 0204-8043 |
| DOI: | 10.2478/v10153-011-0063-0 |
| Popis: | INTRODUCTION: Several immunological methods are used to determine the serum antinuclear antibodies (ANA). Indirect immunofl uorescence assay (IFA) with tissue slices or HEp-2 cells is the standard technique considered the gold standard for their screening. Serumfree McCoy-Plovdiv cell line may also be used as substrate for IFA. Another method for detection of total and specifi c ANA is the enzyme-linked immunosorbent assay (ELISA). Immunoblot is also applied in specifi c ANA confi rmation. The AIM of the current study was to determine and propose a justifi ed immunological approach for identifi cation of clinically signifi cant ANA by comparing the screening tests - ANA-IFA on serum-free McCoy-Plovdiv cell substrate with ELISA for total ANA, and confi rmative methods for specifi c ANA-ELISA with immunoblot. MATERIALS AND METHODS: Serum samples from 38 patients screened for total ANA by ELISA (Trinity Biotech, NY, USA) and IFA-ANA with МcCoy-Plovdiv cell line, were included in the study. Positive samples were confi rmed by immunoblot (Orgentec Diagnostika, Germany) and ELISA for specifi city of confi rmed ANA. RESULTS: No significant difference (Р > 0.05) and very good agreement were found between the two screening tests. Very good agreement for specifi c antibodies against SS-A, SS-B, dsDNA, moderate for anti-Sm and anti-Sm/RNP and fair for anti-histone/nucleosomal antibodies was found between confi rmative methods. No agreement was found for anti-Scl-70 antibodies. CONCLUSION: IFA-ANA with serum-free МcCoy-Plovdiv cell line and screening ELISA may be recommended for determination of total ANA, and immunoblot and ELISA - for confi rmation and identifi cation of specifi c ANA. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|