Abstract:

【Objective】 To produce a visual protein chip that can detect antibodies against avian influenza virus, newcastle disease virus, infectious bronchitis virus and infectious bursal disease virus simultaneously. 【Method】 To prepare protein chips, the four kinds of virus protein were purified, respectively, by ultracentrifugation and sucrose density gradient centrifugalization method , and then they were spotted on a aldehyde group-modified glass slides with a proper concentration. The four kinds of virus protein on the protein chip were reacted with the corresponding positive serum against these viruses, and then were hybridized with nano-gold labeled second antibody. The result was obtained after silver staining. The platform of a visual protein chip to differentiate antibodies of four poultry diseases was constructed. Furthermore, specificity and the correlation of the gray value scale with the antibody concentration of this visual protein chip were studied. Finally, the visual protein chip was used to test 18 field serum specimens. 【Result】 The results showed that after the conditions were optimized, the probe of this protein chip could be hybridized with corresponding antibodies specifically and strong signal was presented without cross-hybridization. The gray value scale showed linear in the concentration range of antibodies at 50-300 μg?ml-1. This protein chip showed good sensibility and sensitivity compared with traditional method when they were used to detect 18 serum samples simultaneouly. 【Conclusion】 The protein chip reported here can be used to differentiate antibodies of four poultry diseases.

Key words: visual, protein chip, antibodies of poultry diseases, differentiation