Match-paired monoclonal antibodies against major royal jelly protein 4 (MRJP4) are secreted by hybridoma cell lines having microbial deposit numbers of CGMCC No. 17294 and CGMCC No. 17295, which are used in an ELISA kit and a colloidal gold immunoassay strip for detecting the MRJP4. The positive and MRJP4-sepcific cell lines are obtained by cell fusion using an antigen of MRJP4 recombinant protein and a cross-reaction with other major royal jelly proteins. The MRJP4 recombinant protein is used as an antigen to obtain several positive and cell lines by cell fusion and twoMRJP4-specific fusion cell lines are obtained by a cross-reaction with other major royal jelly proteins. Primary screening of a matched antibody pair is performed according to antibody pairing for recognizing different epitopes.