Toll-like receptor (TLR) genes have recently been used to assess the adaptive genetic diversity under natural selection. Here, we newly developed a set of primer pairs for amplifying TLR genes (TLR1LB, TLR2A, TLR3, TLR4, TLR5, TLR7 and TLR15) in the vulnerable Chinese egret (Egretta eulophotes). The PCR products, ranged between 407 and 988 bp (an average of 762 bp), were successfully amplified by using this primer set. Sequence analyses of all these TLR genes indicated that there were 22–75 variable sites and 20–77 single nucleotide polymorphisms. This primer set was also confirmed useful for nine other ardeid species, including four members in ardeinae and five in botaurinae. The versatility of this primer set will provide a useful groundwork for the further studies on the TLR conservation genetics of this vulnerable egret and other ardeid birds.