Introduction: Several immunological methods are used to determine the serum antinuclear antibodies (ANA). Indirect immunofluorescence assay (IFA) with tissue slices or HEp-2 cells is the standard technique considered the gold standard for their screening. Serum-free McCoy-Plovdiv cell line may also be used as substrate for IFA. Another method for detection of total and specific ANA is the enzyme-linked immunosorbent assay (ELISA). Immunoblot is also applied in specific ANA confirmation. The aim of the current study was to determine and propose a justified immunological approach for identification of clinically significant ANA by comparing the screening tests - ANA-IFA on serum-free McCoy-Plovdiv cell substrate with ELISA for total ANA, and confirmative methods for specific ANA-ELISA with immunoblot.
Materials And Methods: Serum samples from 38 patients screened for totalANA by ELISA (Trinity Biotech, NY, USA) and IFA-ANA with McCoy-Plovdiv cell line, were included in the study. Positive samples were confirmed by immunoblot (Orgentec Diagnostika, Germany) and ELISA for specificity of confirmed ANA.
Results: No significant difference (P > 0.05) and very good agreement were found between the two screening tests. Very good agreement for specific 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 confirmative methods. No agreement was found for anti-Scl-70 antibodies.
Conclusion: IFA-ANA with serum-free McCoy-Plovdiv cell line and screening ELISA may be recommended for determination of total ANA, and immunoblot and ELISA- for confirmation and identification of specific ANA.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.2478/v10153-011-0063-0 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Cytotoxic effect of kernel extract on HCT116 and A2058 cancer cell lines.
Heliyon
September 2020
Department of Chemical Sciences, Faculty of Pharmacy, Medical University of Plovdiv, 15A Vasil Aprilov Blvd, Plovdiv, 4000, Bulgaria.
While the pharmacology of leaf extract has been studied extensively, little is known about the pharmacological potential of seeds, although they contain similar active ingredients that are responsible for the therapeutic effects of the leaf extract. In this study we used 70%-methanol kernel extract, quantified its bioactive constituents and tested their cytotoxic effect on two cancer cell lines, A2058 and HCT116, and the non-tumor cell line McCoy-Plovdiv. We studied the biological effect of the extract by real-time analysis in the xCELLigence system, WST-1 assay and LIVE/DEAD viability assay.
View Article and Find Full Text PDFInvestigation Of Chromium-Cobalt Coated Scaffolds On Cell Cultures In Vitro.
Folia Med (Plovdiv)
December 2015
Department of Medical Biology, Medical Faculty, Medical University, Plovdiv, Bulgaria
Introduction: The use of diverse materials for medical purposes is continuously expanding. The modification of materials which are routinely applied in medical practice as well as the development and introduction of new materials require studies on their biological activity. The first steps in this process are the preliminary short-term screening tests for cytotoxicity and biocompatibility performed on cell cultures.
View Article and Find Full Text PDFMicromeritic procedures in assessing antinuclear antibody patterns in immunofluorescent assay.
Folia Med (Plovdiv)
January 2015
Institute for Nuclear Research and Nuclear Energy, Bulgarian Academy of Sciences, Sofi a, Bulgaria
Aim: The aim of this study was to introduce a micromeritic procedure (a statistical approach for small objects) in indirect immunofluorescence assay (IFA) to find objective quantitative parameters of antinuclear antibody (ANA) patterns which could support a diagnosis of auto-immune diseases.
Materials And Methods: Sera of patients with systemic autoimmune diseases, McCoy-Plovdiv serum-free cell line, goat anti-human immunoglobulin-G FITC-conjugate, fluorescent microscope, computer-assisted digital image processing, analysis using a micromeritic procedure, ANOVA.
Results: Three ANA fluorescent patterns (homogeneous, rim and speckled) were analyzed by the micromeritic procedure.
A comparative study of immunological methods for determination of serum antinuclear antibodies.
Folia Med (Plovdiv)
July 2012
Research Immunology Center Medical University, Plovdiv, Bulgaria.
Introduction: Several immunological methods are used to determine the serum antinuclear antibodies (ANA). Indirect immunofluorescence assay (IFA) with tissue slices or HEp-2 cells is the standard technique considered the gold standard for their screening. Serum-free McCoy-Plovdiv cell line may also be used as substrate for IFA.
View Article and Find Full Text PDFGuidelines for cultivation and preservation of the serum-free cell line McCoy-Plovdiv.
Cytotechnology
September 2003
Department of Developmental Biology, The University of Plovdiv, 24, Tzar Assen Str., 4000 Plovdiv, Bulgaria(e-mail,
In this paper, we offer detailed information concerning manipulations with the novel serum-free cell line McCoy-Plovdiv. Guidelines for procedures as trypsinization of the monolayer, subculturing, as well as freezing and thawing conditions are proposed. Our results give us grounds to assume that this is a cell line entirely serum-independent at any step of the process of culturing and preservation.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!