Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Confirmation of Male Specific Fetal Free RNA in Maternal Plasma and Comparison of Accuracy on the Sex Determination using Real-time PCR Method in Korean Native Cattle

  • Lee, Sang-Ho (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University) ;
  • Park, Chul-Ho (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University) ;
  • Park, Jun-Tae (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University) ;
  • Park, Sang-Guk (Rural Development Administration, National Institute of Animal Science) ;
  • Lee, Jin-A (Laboratory of Veterinary Infectious Disease, College of Veterinary Medicine, Chonnam National University) ;
  • Suh, Guk-Hyun (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University) ;
  • Oh, Ki-Seok (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University) ;
  • Son, Chang-Ho (Laboratory of Veterinary Obstetrics, College of Veterinary Medicine, Chonnam National University)
  • Received : 2013.12.06
  • Accepted : 2013.12.18
  • Published : 2013.12.31
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Abstract

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

Keywords

References

  1. Ali A. 2004. Effect of gestational age and fetal position on the possibility and accuracy of ultrasonographic fetal gender determination in dairy cattle. Reprod. Dom. Anim. 39: 190-194. https://doi.org/10.1111/j.1439-0531.2004.00502.x
  2. Bianchi DW. 2004. Circulating fetal DNA: its origin and diagnostic potential-a review. Placenta 25: S93-101. https://doi.org/10.1016/j.placenta.2004.01.005
  3. da Cruz AS, Silva DC, Costa EO, De M-Jr P, da Silva CC, Silva DM and da Cruz AD. 2012. Cattle fetal sex determination by polymerase chain reaction using DNA isolated from maternal plasma. Anim. Reprod. Sci. 131: 49-53. https://doi.org/10.1016/j.anireprosci.2012.02.004
  4. de Leon PM, Campos VF, Dellagostin OA, Deschamps JC, Seixas FK and Collares T. 2012. Equine fetal sex determination using circulating cell-free fetal DNA (ccffDNA). Theriogenology 77: 694-698. https://doi.org/10.1016/j.theriogenology.2011.09.005
  5. Finning KM, Chitty LS. 2008. Non-invasive fetal sex determination: impact on clinical practice. Semin. Fetal. Neonatal. Med. 13: 69-75. https://doi.org/10.1016/j.siny.2007.12.007
  6. Hamilton CK, Combe A, Caudle J, Ashkar FA, Macaulay AD, Blondin P and King WA. 2012. A novel approach to sexing bovine blastocysts using male-specific gene expression. Theriogenology 77: 1587-1596. https://doi.org/10.1016/j.theriogenology.2011.11.027
  7. Heid CA, Stevens J, Livak KJ and Williams PM. 1996. Real time quantitative PCR. Genome Res. 6: 986-994. https://doi.org/10.1101/gr.6.10.986
  8. Jimenez DF and Tarantal AF. 2003. Fetal gender determination in early first trimester pregnancies of rhesus monkeys (Macaca mulatta) by fluorescent PCR analysis of maternal serum. J. Med. Primatol. 32: 315-319. https://doi.org/10.1046/j.1600-0684.2003.00041.x
  9. Kadivar A, Hassanpour H, Mirshokraei P, Azari M, Gholamhosseini K and Karami A. 2013. Detection and quantification of cell-free fetal DNA in ovine maternal plasma; use it to predict fetal sex. Theriogenology 79: 995-1000. https://doi.org/10.1016/j.theriogenology.2013.01.027
  10. Khorram Khorshid HR, Zargari M, Sadeghi MR, Edallatkhah H, Shahhosseiny MH and Kamali K. 2013. Early fetal gender determination using real-time PCR analysis of cell-free fetal DNA during 6th-10th weeks of gestation. Acta. Med. Iran. 51: 209-214.
  11. Lo YMD, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CWG. 1997. Presence of fetal DNA in maternal plasma and serum. Lancet 350: 485-487. https://doi.org/10.1016/S0140-6736(97)02174-0
  12. Lo YMD, Tein MSC, Lau TK, Haines CJ, Leung TN, Poon PMK. 1998. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. Am. J. Hum Genet. 62: 768-775. https://doi.org/10.1086/301800
  13. Poon LL, Leung TN, Lau TK and Lo YM. 2000. Presence of fetal RNA in maternal plasma. Clin. Chem. 46:1832-1834.
  14. Quintela LA, Barrio M, Pena AI, Becerra JJ, Cainzos J, Herra don PG and Diaz C. 2012. Use of ultrasound in the reproductive management of dairy cattle. Reprod. Domest. Anim. 47: 34-44. https://doi.org/10.1111/j.1439-0531.2012.02032.x
  15. Revil T, Gaffney D, Dias C, Majewski J and Jerome-Majewska LA. 2010. Alternative splicing is frequent during early embryonic development in mouse. BMC Genomics. 11:399. https://doi.org/10.1186/1471-2164-11-399
  16. Shea, BF. 1999. Determining the sex of bovine embryos using polymerase chain reaction results: a six year retrospective study. Theriogenology 51: 841-845. https://doi.org/10.1016/S0093-691X(99)00030-8
  17. Wooding FBP. 1992. Current topic: the synepitheliochorial placenta of ruminants: binucleate cell fusions and hormone production. Placenta 13: 101-113. https://doi.org/10.1016/0143-4004(92)90025-O