Detection of anti-HAY antibody with dot immunogold filtration assay

• Published in: World journal of gastroenterology : WJG 2003, Vol.9 (7), p.1508-1511
• Main Author: Zhong-JunShao De-ZhongXu Yong-PingYan Jing-HuaLi Jing-XiaZhang Zhi-YingZhang Bo-RongPan
• Format: Article
• Language: English
• Subjects: 免疫球蛋白G; 免疫球蛋白M; 抗甲肝病毒抗体; 聚合酶链反应; 酶链免疫吸附实验
• ISSN: 1007-9327; 2219-2840

AIM: To establish a rapid, sensitive and specific immunogold assay for detection of hepatitis A virus infection. METHODS: 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. RESULTS: 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-EUSA (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. CONCLUSION: This assay can detect anti-HAY 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.