The coronavirus disease 19 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and the outbreak has rapidly spread worldwide causing a global severe pneumonia pandemic. Several animals have been proven to be affected by the SARS-CoV-2, posing an enormous threat to humans. However, to confirm the intermediate hosts in the epidemic regions, it is unrealistic to detect its presence in each and every animal one by one. An active transmembrane serine protease 2 (TMPRSS2) which functions in priming of S-protein of SARS-CoV-2 is one of the most important prerequisites for an animal to be an intermediate host, but the utilizing capability of TMPRSS2 for non-human animals is largely unknown. Here, we systematically predicted the ability of TMPRSS2 to prime S-protein of SARS-CoV-2 in 164 vertebrates by analyzing the phylogenetic clustering and possible functional changes induced by amino acid variation in TMPRSS2. Mammals were predicted to be the most likely animals to effectively use the TMPRSS2 for priming S-protein of SARS-CoV-2. Especially, we predicted the possible effective utilizing ability of TMPRSS2 in rats and pigs, which are reported non-susceptible to SARS-CoV-2. Given the wide distribution and the close relationships among human, pig and rat, a periodic and careful monitoring is required for pigs and rats to prevent the possible SARS-CoV-2 transmission to human.