The invention relates to a cynoglossus semilaevis gunther sex-linked microsatellite marker and genetics sex testing method comprising a PCR method and a PCR testing method, wherein the PCR method is used for screening and testing of a cynoglossus semilaevis gunther sex-linked microsatellite marker, clone of female peculiar allelic gene, sequence analysis, peculiar primer design and cynoglossus semilaevis gunther genetics sex; and the PCR testing method is used for creating the cynoglossus semilaevis gunther genetics sex according to the obtained cynoglossus semilaevis gunther sex-linked microsatellite marker primer and female peculiar allelic gene. The invention ensures that the cynoglossus semilaevis gunther sex-linked microsatellite marker and the female peculiar allelic gene are screened for the first time, establishes the PCR technique tested by the cynoglossus semilaevis gunther genetics sex, has the characteristics of high efficiency, accuracy and stability, has important application value in the cynoglossus semilaevis gunther sex control and unisexuality seed production, and also has important application prospect in the sex determination gene research by cloning the cynoglossus semilaevis gunther.