Published online Jul 15, 2003. doi: 10.3748/wjg.v9.i7.1508
Revised: March 24, 2003
Accepted: April 3, 2003
Published online: July 15, 2003
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-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.
CONCLUSION: This assay can detect anti-HAV IgM and IgG simultaneously, and be done within 3 min. The simplicity, rapidity and specificity of the assay were useful for screening and epidemiological study.