SRY-negative XX sex reversal is an inherited or sporadically occurring disorder, where testis development appears in the absence of the SRY gene. Although the molecular background of this intersexuality syndrome in pigs is unknown, it was proposed that familial cases might be inherited as a single autosomal recessive trait. Because DMRT1 (Doublesex and Mab-3 related transcription factor 1) is an autosomal locus in pig (SSC1q21), shows sexually dimorphic expression in swine gonads and has strong significance in vertebrate testis development, the molecular analysis of this gene was performed in previously reported three intersexes (38,XX, SRY-negative), the progeny of a single boar from a Polish farm. The first two exons encoding functional DM (double sex and mab-3) domain and the promoter region (the 5'flanking sequence) (altogether 3894 bp) were sequenced and compared with male and female control pigs (n = 16) and with publicly available sequences. Three different polymorphisms were found in the coding region, one Indel type polymorphism (DNA 142_144indelAGC) causing a deletion of an amino acid (protein S47_G48indelS) and two silent SNPs (DNA G432A and G492A). The promoter region seems to be highly polymorphic, since 17 SNPs and 5 indels were detected. However, the sequences of control males and females were concordant with those of the intersexes. These results indicate that DMRT1 is an unlikely candidate gene for SRY-negative XX sex reversal in pig.