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Evaluation of the efficiency of TaqMan duplex real-time PCR assay for non-invasive pre-natal assessment of foetal sex in equine.
Abstract: Accurate diagnosis of foetal sex in pregnant mare is helpful for many breeders, both for private or commercial purposes. In this study, in order to pre-natal foetal sexing in equine, we used TaqMan duplex real-time PCR to detect the specific regions of SRY and TSPY genes on extracted cell-free foetal DNA from maternal blood. Peripheral blood samples from 50 pregnant Arabian mares with singleton foetuses were collected. Cell-free foetal DNA was extracted from maternal plasma, and duplex real-time PCR assays were performed with TaqMan probes and primers. Amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as control of DNA extraction procedure. From the 50 sampled mares, 28 cases had female and 22 mares had male foetuses. The final results for 46 samples were conclusive, and from them, 43 cases were predicted correctly. Sensitivity, specificity and accuracy of the test were 90.48%, 96% and 93.48%, respectively. In conclusion, a TaqMan duplex real-time PCR was set up to pre-natal detection of foetal sex in equine. The method was fast and decreased the false-positive and false-negative results. The technique can be used as a routine procedure in farms by collecting only a blood sample.
© 2020 Wiley-VCH GmbH.
Publication Date: 2020-12-14 PubMed ID: 32978850DOI: 10.1111/rda.13831Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research article reports on the successful use of TaqMan duplex real-time PCR, a form of gene detection, to accurately determine the sex of unborn horses. The technique is based on genetic markers in the blood of pregnant mares and could be a beneficial tool for horse breeding.
Research Purpose
The purpose of the study was to develop a non-invasive, quick, and precise method to determine the sex of unborn foetuses in horses. The researchers aimed to use the TaqMan duplex real-time PCR technique, which utilizes genetic markers to identify male or female foetuses.
Methodology
- The study involved 50 pregnant Arabian mares, each pregnant with a single foetus.
- Peripheral blood samples were taken from these mares, and cell-free foetal DNA was extracted from the maternal plasma.
- Duplex real-time PCR assays were then performed on this DNA using TaqMan probes and primers that targeted specific regions of the SRY and TSPY genes, which are indicative of male sex.
- The GAPDH gene was also amplified as a control measure to ensure the accuracy of the DNA extraction procedure.
Results
- Out of the 50 mares, 28 were found to be pregnant with female foetuses and 22 with male foetuses.
- The results were conclusive for 46 of the samples, and from these, 43 were accurately predicted.
- The test demonstrated a sensitivity of 90.48%, a specificity of 96%, and an accuracy of 93.48%.
Conclusions
- Overall, the study concluded that TaqMan duplex real-time PCR is a reliable method for non-invasively determining foetal sex in horses.
- Beyond its effectiveness, the technique exhibited rapid results and lowered the instance of false-positive and false-negative findings.
- As the procedure only involves a simple blood sample collection, it could be integrated seamlessly into routine farming operations and greatly assist breeders.
Cite This Article
APA
Kadivar A, Rashidzadeh H, Davoodian N, Nazari H, Dehghani Tafti R, Heidari Khoei H, Seidi Samani H, Modaresi J, Ahmadi E.
(2020).
Evaluation of the efficiency of TaqMan duplex real-time PCR assay for non-invasive pre-natal assessment of foetal sex in equine.
Reprod Domest Anim, 56(2), 287-291.
https://doi.org/10.1111/rda.13831 Publication
Researcher Affiliations
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
- Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Department of Clinical Science, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.
- Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
MeSH Terms
- Animals
- Cell-Free Nucleic Acids / blood
- Female
- Fetus
- Genes, sry
- Genetic Testing / veterinary
- Horses
- Male
- Pregnancy
- Real-Time Polymerase Chain Reaction / methods
- Real-Time Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- Sex Determination Analysis / methods
- Sex Determination Analysis / veterinary
Grant Funding
- 97GRN1M1576 / Shahrekord University
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