Shao Zhong-Jun, Xu De-Zhong, Yan Yong-Ping, Li Jing-Hua, Zhang Jing-Xia, Zhang Zhi-Ying, Pan Bo-Rong
Department of Epidemiology, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
World J Gastroenterol. 2003 Jul;9(7):1508-11. doi: 10.3748/wjg.v9.i7.1508.
To establish a rapid, sensitive and specific immunogold assay for detection of hepatitis A virus infection.
Rabbit monoclonal antibodies to anti-human IgM and IgG (Dako) were dotted on a nitrocellulose membrane (NCM) respectively to capture the human sera IgM and IgG. Then the captured antibodies would conjugate to HAV antigen, which was revealed by mouse anti-HAV IgG conjugated to gold particles. Final results were assessed by blind method.
Sera from 96 patients with acute hepatitis were used for our study. Compared with well-recognized standard (Abbott Laboratory, USA), the sensitivity and specificity of IgM-DIGFA (self-made) were 91.3 % (42/46) and 96.0 % (48/50), and those of IgM-ELISA (Kehua, Shanghai) were 97.8 % (45/46) and 100.0 % (50/50). The identical results were produced from the study with reagents at different conditions, and the study was repeated in 15 negative sera and 10 positive sera. The serum anti-HAV IgG was tested with DIGFA at the same time. In comparison with ELISA, the sensitivity and specificity of DIGFA for IgG anti-HAV were 87.2 % (41/47) and 91.8 % (45/49), respectively.
This assay can detect anti-HAV IgM and IgG simultaneously, and be done within 3 minutes. The simplicity, rapidity and specificity of the assay were useful for screening and epidemiological study.
建立一种快速、灵敏且特异的免疫金测定法用于检测甲型肝炎病毒感染。
将抗人IgM和IgG的兔单克隆抗体（Dako公司）分别点样于硝酸纤维素膜（NCM）上，以捕获人血清中的IgM和IgG。然后捕获的抗体与甲型肝炎病毒抗原结合，该抗原由与金颗粒偶联的小鼠抗甲型肝炎病毒IgG显示。最终结果采用盲法评估。
96例急性肝炎患者的血清用于本研究。与公认的标准方法（美国雅培实验室）相比，自制的IgM - DIGFA的敏感性和特异性分别为91.3%（42/46）和96.0%（48/50），而上海科华的IgM - ELISA的敏感性和特异性分别为97.8%（45/46）和100.0%（50/50）。在不同条件下使用试剂进行的研究产生了相同的结果，并且在15份阴性血清和10份阳性血清中重复了该研究。同时用DIGFA检测血清抗甲型肝炎病毒IgG。与ELISA相比，DIGFA检测抗甲型肝炎病毒IgG的敏感性和特异性分别为87.2%（41/47）和91.8%（45/49）。
该测定法可同时检测抗甲型肝炎病毒IgM和IgG，且3分钟内即可完成。该测定法的简便性、快速性和特异性对筛查及流行病学研究有用。
