title: Equine fetal sex determination using circulating cell-free fetal DNA (ccffDNA).
abstract: In this study, polymerase chain reaction (PCR) reamplification of the first PCR product (2nd-PCR) and a qPCR assay were used to detect the sex determining region Y (SRY) gene from circulating cell-free fetal DNA (ccffDNA) in blood plasma of pregnant mares to determine fetal sex. The ccffDNA was isolated from plasma of 20 Thoroughbred mares (5-13 y old) in the final 3 mo of pregnancy (fetal sex was verified after foaling). For controls, plasma from two non-pregnant mares and two virgin mares were used, in addition to the non-template control. The 182 bp nucleotide sequence corresponding to the SRY-PCR product was confirmed by DNA sequencing. Based on SRY/PCR, 8 of 11 male and 9 of 9 female fetuses were correctly identified, resulting in a sensitivity of 72.7% (for male fetuses) and an overall accuracy of 85%. Furthermore, using SRY/2nd-PCR and qPCR techniques, sensitivity and accuracy were 90.9 and 95%, respectively. In conclusion, this study is apparently the first report of fetal sex determination in mares using ccffDNA.
authors: Laboratório de Embriologia Molecular e Transgênese, Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Priscila Marques Moura
authors: PM
authors: de Leon
authors: Vinicius Farias
authors: VF
authors: Campos
authors: Odir Antônio
authors: OA
authors: Dellagostin
authors: João Carlos
authors: JC
authors: Deschamps
authors: Fabiana Kömmling
authors: FK
authors: Seixas
authors: Tiago
authors: T
authors: Collares
mesh_terms: D000818:Animals; D002474:Cell-Free System; D004247:DNA / Q000235:genetics*; D005260:Female; D024282:Genes, sry; D006736:Horses / Q000097:blood* / Q000235:genetics*; D008297:Male; D011247:Pregnancy; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods / Q000662:veterinary*
chemical_list: D004247:DNA