Akatsuka T, Tohmatsu J, Yoshihara N, Katsuhara N, Okamoto T, Shikata T, Odaka T
J Med Virol. 1986 Sep;20(1):33-42. doi: 10.1002/jmv.1890200106.
The antibody to AN6520 antigen, which was isolated from the liver of a patient with non-A, non-B hepatitis (NANBH), has been detected frequently in convalescent sera from patients with NANBH by the passive hemagglutination (PHA) test. In a further study, we established hybridoma cells secreting antibodies against AN6520 antigen and obtained ascitic fluids with PHA titers ranging from 1:10(5) to 1:10(7). In immunodiffusion with AN6520 antigen, all monoclonal antibodies were found to form an identical precipitin line. These lines were also identical to those formed by rabbit antiserum against AN6520 antigen and by convalescent sera from patients with NANBH. With one of the monoclonal antibodies, 1-F12, solid-phase radioimmunoassay (SP-RIA) for detecting AN6520 antigen was developed as well as blocking RIA for anti-AN6520 antibody detection. The antigen assay was 50 times more sensitive than the reverse passive hemagglutination (R-PHA) test, with a sensitivity threshold of the 1 ng/ml of antigen solution; the antibody assay was 10 times more sensitive than PHA. The results with this blocking RIA were mostly in agreement with the data obtained by PHA. Furthermore, the antigen in human sera, which had never been detected by R-PHA test, could be detected by SP-RIA.
从一名非甲非乙型肝炎（NANBH）患者肝脏中分离出的抗AN6520抗原抗体，通过被动血凝试验（PHA）在NANBH患者的恢复期血清中经常被检测到。在进一步的研究中，我们建立了分泌抗AN6520抗原抗体的杂交瘤细胞，并获得了PHA效价范围为1:10(5)至1:10(7)的腹水。在与AN6520抗原的免疫扩散试验中，发现所有单克隆抗体都形成了相同的沉淀线。这些沉淀线也与兔抗AN6520抗原血清和NANBH患者恢复期血清形成的沉淀线相同。利用其中一种单克隆抗体1-F12，开发了用于检测AN6520抗原的固相放射免疫分析（SP-RIA）以及用于检测抗AN6520抗体的阻断RIA。抗原检测比反向被动血凝试验（R-PHA）灵敏50倍，抗原溶液的灵敏度阈值为1 ng/ml；抗体检测比PHA灵敏10倍。这种阻断RIA的结果与PHA获得的数据大多一致。此外，R-PHA试验从未检测到的人血清中的抗原，可通过SP-RIA检测到。
