The invention provides a protein chip for detecting respiratory virus antigens as well as a kit and a preparation method thereof. The protein chip for detecting the respiratory virus antigens comprises a glass matrix vector and a quality control protein probe fixed in a protein quality comparison hole in the glass matrix vector; quality control protein comprises an adenovirus antigen, a respiratory syncytial virus antigen, influenza A and B antigens, a parainfluenza virus antigen, a SARS virus antigen, an H5N1 antigen and an H1N1-a antigen. Through adoption of the technical scheme, the diagnosis of simultaneous detection of eight types or more types of viral infection can be realized through one experiment, so that the efficiency in diagnosis of respiratory viral infectious diseases is improved, anti-bioterrorism screening is assisted, and a purpose of efficient, time-saving and high-flux diagnosis is realized.