Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production
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2018
Authors
Ristanić, MarkoStanišić, Ljubodrag
Maletić, Milan
Glavinić, Uroš
Drašković, Vladimir
Aleksić, Nevenka
Stanimirović, Zoran
Article (Published version)
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Contents Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender-specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell-free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300-600bp) in maternal circulation. The aim of this study was to assess this non-invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non-pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three ...different methods: using tubes with silicone membranes, a single-tube extraction without silicone membranes and phenol-chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real-time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single-tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real-time PCR test sensitivity in Group I was 90% and in Group II 91.6%.
Keywords:
bovine / ccffDNA / foetal sexing / PCR / real-time PCRSource:
Reproduction in Domestic Animals, 2018, 53, 4, 947-954Publisher:
- Wiley, Hoboken
Funding / projects:
- International Atomic Energy AgencyInternational Atomic Energy Agency [20774]
- Molecular genetic and ecophysiological researches on the protection of autochthonous animal genetic resources, sustaining domestic animals’ welfare, health and reproduction, and safe food production (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46002)
DOI: 10.1111/rda.13193
ISSN: 0936-6768
PubMed: 29740884
WoS: 000438515700014
Scopus: 2-s2.0-85046534945
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Institution/Community
Fakultet veterinarske medicineTY - JOUR AU - Ristanić, Marko AU - Stanišić, Ljubodrag AU - Maletić, Milan AU - Glavinić, Uroš AU - Drašković, Vladimir AU - Aleksić, Nevenka AU - Stanimirović, Zoran PY - 2018 UR - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1652 AB - Contents Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender-specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell-free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300-600bp) in maternal circulation. The aim of this study was to assess this non-invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non-pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single-tube extraction without silicone membranes and phenol-chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real-time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single-tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real-time PCR test sensitivity in Group I was 90% and in Group II 91.6%. PB - Wiley, Hoboken T2 - Reproduction in Domestic Animals T1 - Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production VL - 53 IS - 4 SP - 947 EP - 954 DO - 10.1111/rda.13193 ER -
@article{ author = "Ristanić, Marko and Stanišić, Ljubodrag and Maletić, Milan and Glavinić, Uroš and Drašković, Vladimir and Aleksić, Nevenka and Stanimirović, Zoran", year = "2018", abstract = "Contents Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender-specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell-free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300-600bp) in maternal circulation. The aim of this study was to assess this non-invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non-pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single-tube extraction without silicone membranes and phenol-chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real-time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single-tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real-time PCR test sensitivity in Group I was 90% and in Group II 91.6%.", publisher = "Wiley, Hoboken", journal = "Reproduction in Domestic Animals", title = "Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production", volume = "53", number = "4", pages = "947-954", doi = "10.1111/rda.13193" }
Ristanić, M., Stanišić, L., Maletić, M., Glavinić, U., Drašković, V., Aleksić, N.,& Stanimirović, Z.. (2018). Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production. in Reproduction in Domestic Animals Wiley, Hoboken., 53(4), 947-954. https://doi.org/10.1111/rda.13193
Ristanić M, Stanišić L, Maletić M, Glavinić U, Drašković V, Aleksić N, Stanimirović Z. Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production. in Reproduction in Domestic Animals. 2018;53(4):947-954. doi:10.1111/rda.13193 .
Ristanić, Marko, Stanišić, Ljubodrag, Maletić, Milan, Glavinić, Uroš, Drašković, Vladimir, Aleksić, Nevenka, Stanimirović, Zoran, "Bovine foetal sex determinationDifferent DNA extraction and amplification approaches for efficient livestock production" in Reproduction in Domestic Animals, 53, no. 4 (2018):947-954, https://doi.org/10.1111/rda.13193 . .