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AIM:To establish a rapid,sensitive and specific immunogoldassay for detection of hepatitis A virus infection.METHODS:Rabbit monoclonal antibodies to anti-humanIgM and IgG (Dako) were dotted on a nitrocellulosemembrane (NCM) respectively to capture the human seraIgM and IgG.Then the captured antibodies would conjugateto HAV antigen,which was revealed by mouse anti-HAVIgG conjugated to gold particles.Final results were assessedby blind method.RESULTS:Sera from 96 patients with acute hepatitis wereused for our study.Compared with well-recognized standard(Abbott Laboratory,USA),the sensitivity and specificity ofIgM-DIGFA (self-made) were 91.3% (42/46) and 96.0%(48/50),and those of IgM-ELISA (Kehua,Shanghai) were97.8% (45146) and 100.0% (50/50).The identical resultswere produced from the study with reagents at differentconditions,and the study was repeated in 15 negative seraand 10 positive sera.The serum anti-HAV IgG was testedwith DIGFA at the same time.In comparison with ELISA,the sensitivity and specificity of DIGFA for IgG anti-HAV were87.2% (41147) and 91.8% (45149),respectively.CONCLUSION:This assay can detect anti-HAV IgM andIgG simultaneously,and be done within 3 minutes.Thesimplicity,rapidity and specificity of the assay were usefulfor screening and epidemiological study.
AIM: To establish a rapid, sensitive and specific immunogold assay for detection of hepatitis A virus infection. DETHODS: Rabbit monoclonal antibodies to anti-human IgG and IgG (Dako) were dotted on a nitrocellulose membrane (NCM) to capture the human sera IgG and IgG. Then the captured antibodies would conjugate to HAV antigen, which was revealed by mouse anti-HAVIgG conjugated to gold particles. Final results were assessed by blind method .RESULTS: Sera from 96 patients with acute hepatitis wereused for our study. Compared with well-recognized standard ( ELISA (Kehua, Shanghai) were 97.8% (42/46) and 96.0% (48/50), the sensitivity and specificity of IgG-DIGFA (45146) and 100.0% (50/50). The identical resultswere produced from the study with reagents at different conditions, and the study was repeated in 15 negative seraand 10 positive sera. The serum anti-HAV IgG was tested with DIGFA at the same time In comparison with ELISA, the sensit ivity and specificity of DIGFA for IgG anti-HAV were 87.2% (41147) and 91.8% (45149) respectively. CONCLUSION: This assay can detect anti-HAV IgM and IgG simultaneously, and be done within 3 minutes. specificity of the assay were useful for screening and epidemiological study.