The invention discloses a primer group, a kit and a method for detecting six mouse viruses by multiple immunity fluorescence. The primer group is designed by the aid of multiple RT-PCR (reverse transcription-polymerase chain reaction) technologies and Luminex liquid chip technologies and can simultaneously detect six high-infection viruses such as TMEV, MHV, MNV, Reo-3, MVM and MPV of mice. By the aid of the primer group, target amplification products are acquired through specific PCR, the amplification products, fluorescence encoding microspheres and streptavidin-phycoerythrin are hybridized, and different types of viruses are distinguished when a mean fluorescence intensity value is read by a Luminex detector. The method has the advantages of rapidness, high efficiency, high specificity and sensitivity, good repeatability and the like and can be applied to quality monitoring, epidemiological investigation and early warning of test mice. The primer group is good in flexibility, and varieties detecting the viruses can be increased and decreased as required on the basis.