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.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2011-10-13 PubMed ID: 22000028DOI: 10.1016/j.theriogenology.2011.09.005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper centers on a study that used circulating cell-free fetal DNA (ccffDNA) to determine the gender of fetuses in pregnant mares. The sex determining region Y (SRY) gene was identified in the ccffDNA sample via two methods: a qPCR assay and reamplification of the first PCR product (2nd-PCR).
Methodology
- The research team extracted ccffDNA from plasma of 20 thoroughbred mares that are between 5-13 years old and are in the last three months of their pregnancies. The sex of these fetuses was verified after birth.
- The test group was counter checked with a control group that was made up of plasma from two non-pregnant mares and two virgin mares, and a non-template control.
- After applying PCR (Polymerase Chain Reaction), the team isolated a nucleotide sequence of 182 base pairs that matched the SRY-PCR product, confirming it with DNA sequencing.
Results
- Of the thoroughbred mares tested, the SRY/PCR method accurately determined the sex in 9 of the 9 female fetuses and in 8 of the 11 male fetuses. This outcome offers a sensitivity rate of 72.7% for male fetuses and an overall accuracy rate of 85%.
- The SRY/2nd-PCR and qPCR techniques provided increased sensitivity and accuracy, at 90.9% and 95% respectively.
Conclusion
- The researchers concluded that the study is probably the first to report fetal sex determination in mares using ccffDNA.
- The findings suggest that ccffDNA could be potentially valuable in equine breeding programs where knowing the sex of the fetus is important.
Cite This Article
APA
de Leon PM, Campos VF, Dellagostin OA, Deschamps JC, Seixas FK, Collares T.
(2011).
Equine fetal sex determination using circulating cell-free fetal DNA (ccffDNA).
Theriogenology, 77(3), 694-698.
https://doi.org/10.1016/j.theriogenology.2011.09.005 Publication
Researcher Affiliations
- Laboratório de Embriologia Molecular e Transgênese, Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
MeSH Terms
- Animals
- Cell-Free System
- DNA / genetics
- Female
- Genes, sry
- Horses / blood
- Horses / genetics
- Male
- Pregnancy
- Sensitivity and Specificity
- Sex Determination Analysis / methods
- Sex Determination Analysis / veterinary
Citations
This article has been cited 7 times.- Panizzi L, Dittmer KE, Vignes M, Doucet JS, Gedye K, Waterland MR, Rogers CW, Sano H, McIlwraith CW, Riley CB. Plasma and Synovial Fluid Cell-Free DNA Concentrations Following Induction of Osteoarthritis in Horses. Animals (Basel) 2023 Mar 14;13(6).
- Vincze B, Gáspárdy A, Biácsi A, Papp EÁ, Garamvölgyi L, Sós E, Cseh S, Kovács G, Pádár Z, Zenke P. Sex determination using circulating cell-free fetal DNA in small volume of maternal plasma in elephants. Sci Rep 2019 Oct 24;9(1):15254.
- Mhatre M, Adeli S, Norwitz E, Craigo S, Phillippe M, Edlow A. The Effect of Maternal Obesity on Placental Cell-Free DNA Release in a Mouse Model. Reprod Sci 2019 Sep;26(9):1218-1224.
- Phillippe M, Adeli S. Cell-free DNA release by mouse placental explants. PLoS One 2017;12(6):e0178845.
- Divar MR, Sharifiyazdi H, Kafi M. Application of polymerase chain reaction for fetal gender determination using cervical mucous secretions in the cow. Vet Res Commun 2012 Dec;36(4):215-20.
- de Lara NSS, Weiss RR, Oba E, Kozicki LE, Souza FA, Bergstein-Galan TG, Muehlbauer E, Silvestri M, de Lima PHL, Vaz ES. Prediction Values for the Influence of Fetal Sex on Plasma Progesterone Concentration in Crioulo Breed Mares: A Preliminary Investigation. Reprod Domest Anim 2025 Oct;60(10):e70131.
- Kőszegi H, Zorkóczy O, Biácsi A, Sós E, Gáspárdy A, Zenke P. A Generalized Approach to Genetic Sex Detection of Uniparous Mammalian Species, With High Potential in Foetal Sex Determination Using Circulating Cell-Free DNA of Maternal Plasma. Vet Med Sci 2025 May;11(3):e70311.
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