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.

与传统的免疫检测条不同，开发了一种使用灭活细菌抗原代替抗体作为检测线，以山羊抗小鼠 IgG-FITC 作为示踪剂的新型抗原免疫层析荧光条 (AICFS)。AICFS的适用性调查表明，E.coli O157:H7 (D3) 和Acidovorax citrulli (6F) 杂交瘤细胞培养物可以检测到，但副溶血性弧菌(H7, C9) 杂交瘤细胞培养物与间接酶联免疫吸附测定 (ELISA) 相比被遗漏了。测量并比较了四种抗体亲和力常数 (Ka)，AICFS 可适用于高亲和力抗体检测。与传统的间接 ELISA 相比，AICFS 对 D3 细胞培养物、腹水和纯化抗体的敏感性至少高出 2 倍，通过比较解释对 D3 细胞培养物特异的 AICFS 是合规的，除了菌株 ATCC 43895，并且间接ELISA错过了它。更重要的是，AICFS方法得到了各种真实样本的证实，它可以用于抗体的不同场景，包括McAb的制备、抗体的有效使用、高亲和力抗体分泌的杂交瘤辅助制备和筛选。它可能是一种极好的替代方法，对于病原菌高质量抗体检测的间接 ELISA 方法相应处理时间不到 5%。这是首次使用AICFS进行细菌高品质抗体检测及应用在不同样本中的报道，展示了高亲和力抗体分泌型杂交瘤筛查的快速辅助工具和高品质抗体应用的绝佳替代方法。