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Development of a PCR-based method to identify fetal sex during IVF cycles

Published online by Cambridge University Press:  18 September 2024

Atieh Sadat Mousavi ,
Sadegh Amiri Open the ORCID record for Sadegh Amiri [Opens in a new window] ,
Mehdi Mehdizadeh ,
Mehrdad Bakhtiari ,
Jamileh Sadat Mirsanei ,
Fatemeh Nikmard ,
Mahmood Barati  and
Fatemehsadat Amjadi
Atieh Sadat Mousavi
Affiliation:
Cellular and Molecular Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran
Sadegh Amiri
Affiliation:
Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences (IUMS), Tehran, Iran
Mehdi Mehdizadeh
Affiliation:
Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran
Mehrdad Bakhtiari
Affiliation:
Cellular and Molecular Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran
Jamileh Sadat Mirsanei
Affiliation:
Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran
Fatemeh Nikmard
Affiliation:
Laleh IVF Clinic, Laleh Hospital, Tehran, Iran
Mahmood Barati*
Affiliation:
Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran
Fatemehsadat Amjadi*
Affiliation:
Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences, Tehran, Iran Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences (IUMS), Tehran, Iran
*
Corresponding authors: Mahmood Barati; Email: [email protected]; Fatemehsadat Amjadi; Email: [email protected]
Corresponding authors: Mahmood Barati; Email: [email protected]; Fatemehsadat Amjadi; Email: [email protected]
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Abstract

One of the most recognizable cases of preimplantation genetic diagnosis (PGD) is X-linked diseases. Diagnosis of fetal sex is essential for couples who are known to be at risk of some X-linked disorders. The objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting sex chromosomes-specific sequences in spent culture medium and comparing these results to PGD/CGH array results. It may open new window for the development of a non-invasive PGD method. 120 Embryo’s spent media from Day 3 and Day 5 embryos were collected. Modified phenol-chloroform solution was used for DNA extraction from spent media. Sex determination was performed using SRY, TSPY and AMELOGENIN evaluation through quantitative polymerase chain reaction (q-PCR) method. IBM SPSS and MedCalc were used for statistical analyses to compare sex determination of embryos by spent medium with PGD/CGH array results. Culture time was demonstrated to increase the DNA amount among day 5 embryos culture medium samples. Non-invasive PGD by means of spent culture medium gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% for sex determination. Results of sex determination using spent medium by q-PCR were consistent with the results of PGD/CGH array. Improvements in cell-free DNA extraction and PCR amplification procedures provide us an effective method to perform a PGD test without biopsy in the future, especially about X-linked diseases.

Keywords

cfDNAPCRPGDsex determinationX-link diseases
Type
Research Article
Information
Zygote , First View , pp. 1 - 8
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Copyright
© The Author(s), 2024. Published by Cambridge University Press

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Footnotes

†

These two co-first authors contributed equally to this study.

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