A novel antigen immunochromatography fluorometric strip for rapid detection and application of pathogenic bacterial...

Unlike traditional immunoassay strips, a novel antigen immunechromatography fluorometric strip (AICFS) using inactivated bacterial antigen instead of an antibody as a test line and goat anti-mouse IgG-FITC as a tracer was developed. The applicability survey of AICFS indicated that E. coli O157:H7 (D3) and Acidovorax citrulli (6F) hybridoma cell cultures could be detected, but Vibrio parahemolyticus (H7, C9) hybridoma cell cultures were missed compared with the indirect enzyme-linked immunosorbent assay (ELISA). The four antibody affinity constants (Ka) were measured and compared, and AICFS could be suitable for high-affinity antibody detection. Compared with the traditional indirect ELISA, the AICFS sensitivity for D3 cell cultures, ascites, and purified antibodies was at least 2-fold more sensitive, the AICFS specific for D3 cell cultures by comparative interpretation was compliant except for the strain ATCC 43895, and the indirect ELISA missed it. More importantly, the AICFS method was confirmed by various real samples that it could be used in different scenarios regarding the antibody, including McAb preparation, the effective antibody use, and high-affinity antibody-secreted hybridoma auxiliary preparation and screening. It could be an excellent alternative method with less than 5% corresponding processing time for indirect ELISA method for pathogenic bacterial high-quality antibody detection. This is the first report of using AICFS for bacterial high-quality antibody detection and application in different samples, which demonstrates a rapid auxiliary tool for high-affinity antibody secreted-hybridoma screening and an excellent alternative method for high-quality antibody application.