Human ANA IgG ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

Anti-Nudear Antibodies (ANA) are autoantibodies which binds to cellular nuclear antigens incuding ds-DNA, ss-DNA, histones, ribonuceoproteins (RNP) and the SS-A, SS-B, and Sm antigens. ANA ELISA, a sandwich ELISA, provides a rapid semi-quantitative measurement of ANA in serum to further investigate the presence of specific autoantibodies

Contents of Kit

1. Nuclear antigen coated microwell strips (96 wells)
2. ANA Negative Control A, ready-to-use, 1.5 ml
3. ANA Cut-off Control B, ready-to-use, 1.5 ml
4. ANA Positive Control C, ready-to-use, 1.5 ml
5. Sample Diluent (5×), 20 ml (yellow color)
6. Wash buffer (50×), 20 ml
7. Anti-hIgG HRP Conjugate, 15 ml (light red)
8. HRP Substrate Solution (TMB) , 15 ml
9. Stop Solution, 15 ml
10. Complete Instruction Manual

Storage

The microtiter well plate and all other reagents are stable at 2-8°C until the expiration date printed on the label. The whole kit stability is usually 12 months from the date of shipping under appropriate storage conditions.
Do not contaminate the bottles. Withdraw solutions in a separate clean tube or dispensing trays. Any unused solution should be discarded and not returned to the bottle. Do not use HRP substrate solution if this solution is blue. Do not expose these solutions to strong light.

Performance Characteristics

The ELISA kit was compared with a commercially available ELISA assay (94 samples). The assay show relative sensitivity of 93%, specificity (96%) and agreement (95%).

Precision

Statistics for coefficients of variation (CV) were calculated for each of four samples results of 32 determinations in a single run for Intra-Assay precision Run-to-run prec calculated from the results of 3 different runs with 24 determinations of each sample:

General Description

Rheumatoid autoimmune diseases are often associated with the occurrence of autoantibodies against several nuclear or cytoplasmatic antigens. These so-called anti nuclear antigens (ANA) can be divided into three groups:
1. true anti nuclear antigens (ANA): dsDNA, ssDNA, histones, nucleolic RNA and DNP
2. extractable nuclears antigens: Sm (Smith), n-RNP, Scl 70 and PM-1
3. cytoplasmatic antigens: SS-A (Ro)*, SS-B (La)* and Jo-1 SS-A (Ro) and SS-B (La) are co-localized in cytoplasm and nucleus
Inflammatory connective tissue diseases are characterized by idiopathic genesis along with disturbances in terms of cellular and humoral immunity, systemic organ failure and a chronic course of disease. Additionally, connective tissue diseases exhibit overlapping symptomatic features that render an accurate diagnosis difficult. Considering the diversity of mixed connective tissue diseases, such disorders exhibit a common serological characteristic; the presence of anti-nuclear antibodies. These antibodies are directed against parts of the cell nucleus and the cytoplasm, and many rheumatic diseases are characterized by the presence of one or more of these ANAs. Antibodies to doublestranded DNA (dsDNA), single-stranded DNA (ssDNA), histone, nuclear ribonucleoprotein (RNP) and Smith antigen (Sm) are associated with SLE, while antibodies to Sjogren´s Syndrome A (SSA/Ro) and Sjogren´s Syndrome B (SSB/La) can occur in both SLE and Sjogren´s Syndrome (SS). Antibodies to Jo-1 may be observed in polymyositis and dermatomyositis, while antibodies to scleroderma-associated antigen (Scl-70) and centromere can occur in patients with progressive systemic sclerosis (PSS). Anti-histone antibodies are associated with SLE and drug-induced lupus, while anti-RNP antibodies are linked with mixed connective tissue disease (MCTD) and with SLE. Antibodies directed against centromere are associated with CREST syndrome. Although IFA technology was traditionally used to detect autoantibodies in conjunction with HEp2 cells, it is now widely ackncMiedged that ELISA technology offers an excellent alternative.