A protein chip designed to differentiate visually antibodies in chickens which were infected by four different viruses.
This study aimed to develop a visual protein chip that can differentiate the antibodies induced by avian influenza virus, Newcastle disease virus, infectious bronchitis virus and infectious bursal disease virus, simultaneously. Proteins from the four viruses were purified and spotted onto an aldehyde group-modified glass slide at 2mg/ml. After that, the protein chip was reacted with the corresponding positive serum against these viruses, hybridized with a colloidal gold-labeled secondary antibody and visualized by silver staining. A diagnostic protein chip was constructed to differentiate antibodies of four poultry diseases This protein chip showed good sensitivity compared with traditional methods, and it was more than 400 times as sensitive as the agar gel precipitin methods used to detect avian influenza and infectious bursal disease. The protein chip was used to test known serum samples of the four poultry diseases and field serum samples. The results showed that this method could hybridize specifically with the corresponding antibodies with strong signals and without cross-hybridization. In conclusion, this protein chip can be used to differentiate the antibodies induced by the four avian viruses.
Wang
Xiurong
X
Animal Influenza Laboratory of Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin, China. wxr@hvri.ac.cn
Shi
Lin
L
Tao
Qimeng
Q
Bao
Hongmei
H
Wu
Jianan
J
Cai
Dongdong
D
Wang
Fumei
F
Zhao
Yuhui
Y
Tian
Guobin
G
Li
Yanbing
Y
Qao
Chuanling
C
Chen
Hualan
H
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000302:isolation & purification; D002645:Chickens; D001351:Infectious bronchitis virus / Q000276:immunology / Q000302:isolation & purification; D007243:Infectious bursal disease virus / Q000276:immunology / Q000302:isolation & purification; D009980:Influenza A virus / Q000276:immunology / Q000302:isolation & purification; D009522:Newcastle disease virus / Q000276:immunology / Q000302:isolation & purification; D011201:Poultry Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D040081:Protein Array Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000302:isolation & purification; D014773:Virology / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014764:Viral Proteins