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A comparative study of immunological methods for determination of serum antinuclear antibodies

 

Mariana A. Murdjeva12 / Nadezhda P. Ryasheva3 / Marian M. Draganov4 / Lyubomir D. Paunov56
1Research Immunology Center, Medical University, Plovdiv
2 German-Bulgarian Medical Diagnostic Laboratory “Synwest-K”, Plovdiv
3German-Bulgarian Medical Diagnostic Laboratory “Synwest-K”, Plovdiv
4Department of Developmental Biology, Plovdiv University “Paisii Hilendarski”, Bulgaria
5Research Immunology Center, Medical University, Plovdiv
6 German-Bulgarian Medical Diagnostic Laboratory “Synwest-K”, Plovdiv
Correspondence and reprint request to: M. Murdjeva, Research Immunology Center, Medical University, PlovdivE-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.; Mob.: +359 882 038 80715A Vassil Aprilov St, 4002 Plovdiv, Bulgaria;

Citation Information: Folia Medica. Volume 53, Issue 4, Pages 21–27, ISSN (Online) 1314-2143, ISSN (Print) 0204-8043, DOI: 10.2478/v10153-011-0063-0, October 2012
Publication History:
Published Online:
2012-10-24


ABSTRACT

 

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.