Anti-nuclear antibodies (ANA) are a group of antibodies directed against various nuclear and some cytoplasmic antigens. Serological tests for ANA play an important role towards the diagnosis of various autoimmune connective tissue disorders. Although these antibodies were first associated with systemic lupus erythematosus (SLE), the list of implicated diseases has expanded and many rheumatic diseases are characterized by the presence of one or more of these ANAs. For instance, anti-SSA/Ro and anti-SSB/La antibodies are associated with SLE and Sjögren’s Syndrome (SS), anti-dsDNA and anti-Sm antibodies with SLE, anti-RNP antibodies with mixed connective tissue disease (MCTD) and SLE, anti-Scl-70 antibodies with scleroderma (progressive systemic sclerosis (PSSJ), anti-Jo1 with polymyositis and dermatomyositis and anti-centromere antibodies with CREST syndrome. As ANA ELISA test collectively detects, in one well, total ANAs against double stranded DNA (dsDNA), Sm, U1-RNP (68K), SS-A/Ro, SS-B/La, Scl-70, Jo-1, and centromeric antigens, along with sera positive for IFA HEp-2 ANAs, it is not specific indicators of a connective tissue disease. To monitor more specific antibodies, eight different antigens (dsDNA, SmD1, U1-RNA (68K), SS-A/Ro, SS-B/La, Scl-70, Jo-1, and CENP-B) are coated to different wells in a column or strip for the ELISA screen test of eight different autoimmune antibodies once.