Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment
Background Freemartinism generally occurs in female offspring of dizygotic twins in a mixed-sex pregnancy. Most bovine heterosexual twin females are freemartins. However, about 10% of bovine heterosexual twin females are fertile. Farmers mostly cull bovine fertile heterosexual twin females due to th...
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PeerJ Inc.
2018-04-01
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| author | Qinghua Qiu Taoqi Shao Yang He Aziz-Ur-Rahman Muhammad Binghai Cao Huawei Su |
| author_facet | Qinghua Qiu Taoqi Shao Yang He Aziz-Ur-Rahman Muhammad Binghai Cao Huawei Su |
| author_sort | Qinghua Qiu |
| collection | DOAJ |
| description | Background Freemartinism generally occurs in female offspring of dizygotic twins in a mixed-sex pregnancy. Most bovine heterosexual twin females are freemartins. However, about 10% of bovine heterosexual twin females are fertile. Farmers mostly cull bovine fertile heterosexual twin females due to the lack of a practical diagnostic approach. Culling of such animals results in economic and genetic-material losses both for dairy and beef industry. Methods In this study, a comparative test, including qualitative detection of SRY gene by polymerase chain reaction (PCR), quantitative detection of relative content of SRY by real-time quantitative polymerase chain reaction (qPCR), and quantitative detection of H-Y antigen, was performed to establish the most accurate diagnosis for freemartin. Twelve Holstein heterosexual twin females were used in this study, while three normal Holstein bulls and three normal Holstein cows were used as a positive and negative control, respectively. Results Polymerase chain reaction results revealed that SRY gene were absent in three heterosexual twin females and only two of them were verified as fertile in later age. The qPCR results showed that relative content of SRY was more than 14.2% in freemartins and below 0.41% in fertile heterosexual twin females. The H-Y antigen test showed no significant numerical difference between freemartin and fertile heterosexual twin female. Discussion Our results show that relative content of SRY quantified by qPCR is a better detection method for diagnosis of freemartin in Holstein cattle as compare to qualitative detection of SRY gene by PCR or quantitative detection of H-Y antigen. To the authors’ knowledge, this is the first time we applied qPCR to diagnosing freemartin by quantifying SRY gene and got relative SRY content of each freemartin and fertile heterosexual twin female. We concluded that low-level of SRY would not influence fertility of bovine heterosexual twin female. |
| first_indexed | 2024-03-09T05:13:51Z |
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| language | English |
| last_indexed | 2024-03-09T05:13:51Z |
| publishDate | 2018-04-01 |
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| spelling | doaj.art-4600ec7a5bf2489f8454565a746dbe2f2023-12-03T12:47:08ZengPeerJ Inc.PeerJ2167-83592018-04-016e461610.7717/peerj.4616Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experimentQinghua Qiu0Taoqi Shao1Yang He2Aziz-Ur-Rahman Muhammad3Binghai Cao4Huawei Su5State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, ChinaState Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, ChinaState Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, ChinaUniversity of Agriculture Faisalabad, Institute of Animal and Dairy Sciences, Faisalabad, PakistanState Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, ChinaState Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, ChinaBackground Freemartinism generally occurs in female offspring of dizygotic twins in a mixed-sex pregnancy. Most bovine heterosexual twin females are freemartins. However, about 10% of bovine heterosexual twin females are fertile. Farmers mostly cull bovine fertile heterosexual twin females due to the lack of a practical diagnostic approach. Culling of such animals results in economic and genetic-material losses both for dairy and beef industry. Methods In this study, a comparative test, including qualitative detection of SRY gene by polymerase chain reaction (PCR), quantitative detection of relative content of SRY by real-time quantitative polymerase chain reaction (qPCR), and quantitative detection of H-Y antigen, was performed to establish the most accurate diagnosis for freemartin. Twelve Holstein heterosexual twin females were used in this study, while three normal Holstein bulls and three normal Holstein cows were used as a positive and negative control, respectively. Results Polymerase chain reaction results revealed that SRY gene were absent in three heterosexual twin females and only two of them were verified as fertile in later age. The qPCR results showed that relative content of SRY was more than 14.2% in freemartins and below 0.41% in fertile heterosexual twin females. The H-Y antigen test showed no significant numerical difference between freemartin and fertile heterosexual twin female. Discussion Our results show that relative content of SRY quantified by qPCR is a better detection method for diagnosis of freemartin in Holstein cattle as compare to qualitative detection of SRY gene by PCR or quantitative detection of H-Y antigen. To the authors’ knowledge, this is the first time we applied qPCR to diagnosing freemartin by quantifying SRY gene and got relative SRY content of each freemartin and fertile heterosexual twin female. We concluded that low-level of SRY would not influence fertility of bovine heterosexual twin female.https://peerj.com/articles/4616.pdfHeterosexual twin femaleFreemartinH-Y antigenqPCRSRY |
| spellingShingle | Qinghua Qiu Taoqi Shao Yang He Aziz-Ur-Rahman Muhammad Binghai Cao Huawei Su Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment PeerJ Heterosexual twin female Freemartin H-Y antigen qPCR SRY |
| title | Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment |
| title_full | Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment |
| title_fullStr | Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment |
| title_full_unstemmed | Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment |
| title_short | Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifying SRY gene: a comparison experiment |
| title_sort | applying real time quantitative pcr to diagnosis of freemartin in holstein cattle by quantifying sry gene a comparison experiment |
| topic | Heterosexual twin female Freemartin H-Y antigen qPCR SRY |
| url | https://peerj.com/articles/4616.pdf |
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