The invention discloses a microsatellite primer combination for identifying the genetic relationship between eggs of Lophophorus lhuysii in the same bird house and application. The primers comprise CM34, CM104, CM412, CM687, CM1423, CM1959, CM2014, CM2083, CM2269, CM2289, CM3131 and CM3590, and the nucleotide sequences of the primers are sequentially shown as SEQ ID NO.1-24. The primers can be stably amplified in the eggshell DNA of Lophophorus lhuysii, the microsatellite sequence of Lophophorus lhuysii with high polymorphism content and good repeatability is obtained through the 12 pairs of primers, the genotyping technology corresponding to the markers is established, and identification of three genetic relationship conditions of sib, half-sib or non-paternity between the eggs of Lophophorus lhuysii in the same bird house can be realized.