Application of polymerase chain reaction for fetal gender determination using cervical mucous secretions in the cow

Abstract

In the current study we aimed to use PCR to investigate the presence of fetal DNA in the bovine (Bos taurus) cervical secretions and maternal serum, and to assess the effectiveness of this method in fetal gender determination. Pregnant uteri and pre-slaughter maternal blood samples were collected from 21 Holstein Frisian cows in a local abattoir. Overall, 13 male and 8 female fetuses were included in the study. Cervical mucus was sampled at the laboratory. After DNA extraction, the PCR amplified a 280 bp fragment from the X-chromosome and a 217 bp fragment from the Y-chromosome based on a sex-related polymorphism in the amelogenin locus. The presence of fetal Y-chromosome was confirmed in seven out of 13 cervical mucus samples collected from cows with male fetuses. Overall test sensitivity for correct sex determination based on PCR assay on cervical samples was equal to 71.4 ± 2 %. In contrast, no fetal Y-chromosome DNA was detected in maternal serum samples from cows with male fetuses. This is the first report on validating the presence of fetal DNA material in the bovine cervical mucus and its potential usefulness for fetal sexing. Further investigations are needed to maximize the accuracy and evaluate the practicality of this approach.

Effects of kisspeptin-10 on in vitro proliferation and kisspeptin receptor expression in primary epithelial cell cultures isolated from bovine placental cotyledons of fetuses at the first trimester of pregnancy

“…1C, D). The presence of a Y-chromosome-specific sequence in both cell lines confirmed their fetal origin, as reported in previous studies ( [23], literature cited in Ref. 23).…”

“…The presence of a Y-chromosome-specific sequence in both cell lines confirmed their fetal origin, as reported in previous studies ( [23], literature cited in Ref. 23). By gel electrophoresis of PCR products amplified from the two cell lines, two fragments of 280 and 217 bp indicating the presence of X-and Ychromosome-specific sequences were observed (male pattern; Fig.…”

“…As negative control for the presence of Y-chromosome, bovine genomic DNA isolated from ovarian surface epithelial cells was used. The sequence length polymorphisms between bovine amelogenin X and amelogenin Y genes (AMLX/Y) were used as markers for sexing the bovine fetal cells, as previously described in other bovine cell systems [23]. The oligonucleotide sequences of used primers (Primm S.R.L., Milano, Italy) were (forward) × 5 0 -CAGCCAAACCTCCCTCTGC-3 0 and (reverse) 5 0 -CCCGCTTGGTCTTGTCTGTTGC-3 0 .…”

A New Disease Caused by an Unidentified Etiological Agent Affects European Salamanders

“…Despite some similarities with Amphibiocystidium, we failed to detect its DNA and, even though we attempted multiple PCR primers and techniques, we only amplified the DNA of contaminants. The mucus of cysts could inhibit DNA extraction or amplification by reducing their efficiency [43,44], still a modification of the DNA extraction protocol according to [28] did not provide significant improvements. The absence of DNA amplification could be because, even if the cysts may attain conspicuous sizes, the encapsulated cells were quite sparse, possibly limiting the amount of DNA.…”