ANA IFA and Reflex to Immunoblot Test Price in Haripur Road

Test Description

ANA by Indirect Immunofluorescence Assay (IFA) with Reflex to Immunoblot is a two-stage autoimmune diagnostic strategy. In the first stage, ANA IFA is performed on HEp-2 cells (human epithelial cell line) — the gold standard substrate for ANA detection — where patient serum is incubated with cells, followed by fluorescein-conjugated anti-human IgG antibody. The presence of ANA is detected by fluorescence microscopy and the result is reported as a titre (e.g., 1:80, 1:160, 1:320, 1:640, 1:1280) and a fluorescence pattern (homogeneous/diffuse, speckled, centromere, nucleolar, cytoplasmic, rim/peripheral, etc.). Positive ANA IFA with titres ≥1:80 (or ≥1:160 depending on laboratory reference) are considered clinically significant. If the ANA IFA is positive, the assay automatically reflexes to ANA Immunoblot (ENA/ANA Profile) to identify the specific autoantibody (anti-dsDNA, anti-Sm, anti-Ro, anti-La, anti-Scl-70, etc.). This reflexed approach is cost-effective, clinically logical, and ensures that only relevant patients receive the expensive confirmatory panel. ANA patterns guide the differential diagnosis: homogeneous pattern suggests anti-dsDNA or anti-histone; speckled pattern suggests anti-Sm, anti-Ro, anti-La, or anti-RNP; centromere pattern is highly specific for limited SSc; nucleolar pattern suggests anti-Scl-70, anti-fibrillarin, or anti-RNA polymerase III.