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Detection of Antinuclear antibody (ANA) is the first step in the diagnosis of autoimmune connective tissue disorder (CTD). The gold standard Laboratory assay for the detection of antinuclear antibodies (ANA) is indirect immunofluorescence (IIF) performed on HEp-2 cells (cultured human epithelial cell. Presence of antinuclear antibodies (ANA) is reflected by positive immunofluorescence staining, but since antinuclear antibodies are further divided into subtypes, precise identification of these autoantibody subtypes and their specific target antigens is not possible with indirect immunofluorescence. Identification of these specific antibodies provides valuable information in the diagnostic evaluation, prognosis, and monitoring of patients with autoimmune connective tissue disorder (CTD). Much known association of these specific autoantibodies with the indirect immunofluorescence (IIF) staining pattern of ANA in CTD can be found in western literature and is considered as reference guide over the world.
Since Immune response to disease, antibody profile and individual immune status differs from person to person and also from population to population, therefore, the present study has been designed to evaluate the definite association between ANA patterns and specific antibodies in the serum in the Saudi population and to document differences/similarities with other populations. To the best of my knowledge, no such research work or data correlating the autoantibodies and their ANA patterns is found in Saudia Arabia.
In this study, we will analyze serum samples from the eastern province of Saudia, referred to immunology laboratory, providing services to a tertiary health center and teaching hospital (KFUH) for ANA testing by Indirect Immunofluorescence method and samples further processed for identification of the specific antibodies by line immunoassay in that population. Later the two will be correlated with one another to establish any definite link between the two.
A systemic autoimmune response is the hallmark of the autoimmune connective tissue disease (CTD) and is characterized by the presence of antinuclear antibodies (ANA). Indirect immunofluorescence (IIF) on human epithelial cell tumor line (Hep-2 cells) is the reference technique used for the detection of antinuclear antibodies (ANA) and is also used as a screening test. Positive immunofluorescence staining indicates the presence of ANA and resulting staining pattern depends on the location of the target antigen. However, it does not, allows precise identification of these specific antibodies against nuclear antigens.
Identification of specific antibodies is performed by specialized techniques such as enzyme-linked immunosorbent assay (ELISA), Western blotting or line immunoassay [1-3]. Literature review reveals some known associations between a broad spectrum of specific antibodies and each specific autoimmune rheumatic disease entity. Most of these associations have been identified with data obtained from various studies on Western population. It is need of time to recognize that individual response to disease, immunity status and type of antibodies, all, depending on genetic makeup and therefore varies from person to person, population to population and place to place. Hence associations between ANA pattern and specific antibodies are known to us from research on samples of the western population cannot be applied to patients of CTD from some other population.
There is neither any data nor any research work correlating antinuclear antibody (ANA) immunofluorescence patterns with the specific antibody immunoprofile in the Saudi population to date.
The study that most closely resembles ours, reported in 2010, by Sebastian et al . This study reports the results of sera tested for ANA using HEp-2010/ liver biochip and a screening dilution of 1:100. In another study by Slater and Shmerling, ANA was performed on HEp-2 cells at a titer of 1: 40 . In Albania, Sulcebe and Morcka also reported a similar study in 1992 . They observed the results of sera tested for ANA using rat liver substrate and a screening dilution of 1:100.
Indirect Immunofluorescence on HEp-2 cell is the standard approach for detecting ANAs, and the staining patterns reveal the location of the target antigen. These patterns correspond to the presence of autoantibodies against different nuclear antigens [7, 8]. Although some staining patterns strongly suggest distinct antibody specificities, additional tests are required to demonstrate antibody reactivities against specific nuclear and cytoplasmic antigens. Identification of the fine specificity may provide valuable assistance in diagnosis, prognosis, and monitoring of patients suffering from rheumatic connective tissue diseases.
In this study, we will analyze serum samples from the eastern province of Saudia, referred to Immunology laboratory, providing services to a tertiary health center and teaching hospital (KFUH) for ANA testing by Indirect Immunofluorescence method and samples further processed for identification of the specific antibodies by line immunoassay in that population and the two will be correlated with one another to establish any definite link between the two.
Data will be analyzed anonymously. The review of medical records will be performed retrospectively on serological tests (IIF and Line immunoassay) and patient data that are performed as part of routine laboratory work.
Aims: To understand a definite association between ANA Immunofluorescence staining patterns and specific autoantibodies in the serum in the Saudi patients suffering from CTD and to document differences and similarities with other populations.
Expected benefits: Absence of data correlating the autoantibodies and their antinuclear antibody (ANA) patterns with the immunoprofile in the Saudi population represents a serious gap in our knowledge.
Data obtained from this study would, therefore, provide a reference database for the Saudi population. In case a definite correlation is found between the ANA staining patterns and the specific antibodies identified by line immunoassay, one could restrict performing line immunoassays which are expensive and use ANA-IIF fluorescent patterns to predict the presence of autoantibodies to precisely diagnose a CTD. This would reduce the cost of laboratory investigations for the patient and would save the resources of Laboratory and hospital used in the diagnosis of Patients suffering from Rheumatic Connective tissue diseases.
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