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A Low-Cost Efficient Multiplex PCR for Prenatal Sex Determination in Bovine Fetus Using Free Fetal DNA in Maternal Plasma
BACKGROUND: In order to establish a reliable non-invasive method for sex determination in a bovine fetus in a routine setting, the possibility of identifying specific sequence in the fetal X and Y-chromosomes has been evaluated in maternal plasma using conventional multiplex polymerase chain reactio...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260644/ https://www.ncbi.nlm.nih.gov/pubmed/25505511 |
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author | Davoudi, Arash Seighalani, Ramin Aleyasin, Seyed Ahmad Tarang, Alireza Salehi, Abdolreza Salehi Tahmoressi, Farideh |
author_facet | Davoudi, Arash Seighalani, Ramin Aleyasin, Seyed Ahmad Tarang, Alireza Salehi, Abdolreza Salehi Tahmoressi, Farideh |
author_sort | Davoudi, Arash |
collection | PubMed |
description | BACKGROUND: In order to establish a reliable non-invasive method for sex determination in a bovine fetus in a routine setting, the possibility of identifying specific sequence in the fetal X and Y-chromosomes has been evaluated in maternal plasma using conventional multiplex polymerase chain reaction (PCR) analysis. The aim of this study was to provide a rapid and reliable method for sexing bovine fetuses. MATERIALS AND METHODS: In this experimental study, peripheral blood samples were taken from 38 pregnant heifers with 8 to 38 weeks of gestation. DNA template was extracted by phenol-chloroform method from 350 µl maternal plasma. Two primer pairs for bovine amelogenin gene (bAML) and BC1.2 were used to amplify fragments from X and Y chromosomes. A multiplex PCR reaction has been optimized for amplification of 467 bp and 341 bp fragments from X and Y bAML gene and a 190 bp fragment from BC1.2 related to Y chromosome. RESULTS: The 467 bp fragment was observed in all 38 samples. Both 341 and 190 bp fragments were detected only in 24 plasma samples from male calves. The sensitivity and specificity of test were 100% with no false negative or false positive results. CONCLUSION: The results showed that phenol-chloroform method is a simple and suitable method for isolation of fetal DNA in maternal plasma. The multiplex PCR method is an available non-invasive approach which is cost efficient and reliable for sexing bovine fetuses. |
format | Online Article Text |
id | pubmed-4260644 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-42606442014-12-10 A Low-Cost Efficient Multiplex PCR for Prenatal Sex Determination in Bovine Fetus Using Free Fetal DNA in Maternal Plasma Davoudi, Arash Seighalani, Ramin Aleyasin, Seyed Ahmad Tarang, Alireza Salehi, Abdolreza Salehi Tahmoressi, Farideh Int J Fertil Steril Original Article BACKGROUND: In order to establish a reliable non-invasive method for sex determination in a bovine fetus in a routine setting, the possibility of identifying specific sequence in the fetal X and Y-chromosomes has been evaluated in maternal plasma using conventional multiplex polymerase chain reaction (PCR) analysis. The aim of this study was to provide a rapid and reliable method for sexing bovine fetuses. MATERIALS AND METHODS: In this experimental study, peripheral blood samples were taken from 38 pregnant heifers with 8 to 38 weeks of gestation. DNA template was extracted by phenol-chloroform method from 350 µl maternal plasma. Two primer pairs for bovine amelogenin gene (bAML) and BC1.2 were used to amplify fragments from X and Y chromosomes. A multiplex PCR reaction has been optimized for amplification of 467 bp and 341 bp fragments from X and Y bAML gene and a 190 bp fragment from BC1.2 related to Y chromosome. RESULTS: The 467 bp fragment was observed in all 38 samples. Both 341 and 190 bp fragments were detected only in 24 plasma samples from male calves. The sensitivity and specificity of test were 100% with no false negative or false positive results. CONCLUSION: The results showed that phenol-chloroform method is a simple and suitable method for isolation of fetal DNA in maternal plasma. The multiplex PCR method is an available non-invasive approach which is cost efficient and reliable for sexing bovine fetuses. Royan Institute 2012 2012-06-19 /pmc/articles/PMC4260644/ /pubmed/25505511 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Davoudi, Arash Seighalani, Ramin Aleyasin, Seyed Ahmad Tarang, Alireza Salehi, Abdolreza Salehi Tahmoressi, Farideh A Low-Cost Efficient Multiplex PCR for Prenatal Sex Determination in Bovine Fetus Using Free Fetal DNA in Maternal Plasma |
title | A Low-Cost Efficient Multiplex PCR for Prenatal Sex
Determination in Bovine Fetus Using Free Fetal DNA in
Maternal Plasma |
title_full | A Low-Cost Efficient Multiplex PCR for Prenatal Sex
Determination in Bovine Fetus Using Free Fetal DNA in
Maternal Plasma |
title_fullStr | A Low-Cost Efficient Multiplex PCR for Prenatal Sex
Determination in Bovine Fetus Using Free Fetal DNA in
Maternal Plasma |
title_full_unstemmed | A Low-Cost Efficient Multiplex PCR for Prenatal Sex
Determination in Bovine Fetus Using Free Fetal DNA in
Maternal Plasma |
title_short | A Low-Cost Efficient Multiplex PCR for Prenatal Sex
Determination in Bovine Fetus Using Free Fetal DNA in
Maternal Plasma |
title_sort | low-cost efficient multiplex pcr for prenatal sex
determination in bovine fetus using free fetal dna in
maternal plasma |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260644/ https://www.ncbi.nlm.nih.gov/pubmed/25505511 |
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