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Detection of antinuclear antibody by Rapid-Dot-ELISA
SUN De-xu QIN Su-ping* DU Wen-ping
Acta Anatomica Sinica ›› 2015, Vol. 46 ›› Issue (1) : 122-126.
Detection of antinuclear antibody by Rapid-Dot-ELISA
Objective To set up a simple and convenient immunoassay for antinuclear antibody (ANA). Methods The ANA positive sera from 20 systemic lupus erythematosus(SLE) cases and the ANA negative sera from 20 healthy were collected and the 40 sera were tested the ANA with Rapid-Dot-ELISA. In the detection we used different concentrations of antigen, different blocking buffer, different concentrations of enzyme-labeled antibodies and different coloration time to choose the optimum testing conditions. The Rapid-Dot-ELISA, ELISA and immunofluorescence methods were used to test the serum ANA in 190 patients with systemic autoimmune diseases and 50 healthy people. Results The concentration of antigen was 40mg/L to 160mg/L, the blocking buffer containing 1% BSA and 10% to 20% bovine serum, the enzyme labeled antibody concentration were 1∶80 to 1∶320, the coloration time in 3 to 5min were the appropriate testing conditions. Compared R-Dot-ELISA with ELISA，the concordance rate of the result was 95.4％（r=0.907, P<0.05）. Compared R-Dot-ELISA with immunofluorescence method，the concordance rate of the result was 96.7％（r=0.932, P<0.05）. Conclusion The sensitivity and specificity of R-Dot-ELISA are high, and the assay is rapid and simple, so R-Dot-ELISA used to detect serum ANA is valuable for application in clinical diagnosis.
Antinuclear antibody / Systemic autoimmune disease / Rapid-Dot-ELISA / Human
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