Non-invasive Preimplantation Genetic Testing for Aneuploidies Using Cell-free DNA in Spent Culture Media (niPGT-A)

ART Fertility Clinics LLC

Status: Withdrawn

Conditions: Secondary Infertility

Treatments: Diagnostic Test: PGT-A / niPGT-A

Study type: Observational

Funder types: Other

Identifiers

NCT04711239

2010-ABU-011-SJ

Details and patient eligibility

About

Analysis of embryonic cell-free DNA (cfDNA) present in the spent culture media (SCM) is a non-invasive alternative for preimplantation genetic testing for aneuploidies (PGT-A) that avoids the technical challenges and limitations of biopsy. Initial studies investigating this non-invasive PGT-A (niPGT-A) method reported variable concordance between cfDNA in SCM and the trophectoderm sample (~ 30%-86%) and indicated a contribution from both the inner cell mass and trophectoderm to the cfDNA in SCM.

This study aims to evaluate the use of the embryo culture medium as a source of genetic material for PGT-A and validate a niPGT-A protocol using cfDNA in SCM.

Full description

Multiple studies have demonstrated the ability to detect and amplify cfDNA from SCM, at different stages of embryonic development, with varying rates of amplification success. Differences in analytes, timing of SCM collection and the duration of embryo culture within the collected medium, performance of assisted hatching (AH), whole genome amplification methods, comprehensive chromosome screening methods and next generation sequencing (NGS) platforms, bioinformatic analyses and strategies for identifying maternal contamination all contribute to the ultimate performance of niPGT-A.

This study aims to validate a noninvasive PGT-A (niPGT-A) method utilizing cfDNA released from the human blastocyst into the SCM.

Patients undergoing a fertility treatment with PGT-A due to secondary infertility will be recruited. On day 6 post fertilization, SCM will be collected prior to blastocyst biopsy. The SCM is normally discarded at this stage. Trophectoderm biopsy and sample collection will follow the IVF laboratory's standard practices for clinical PGT-A.

Three aneuploidy screening kits, relying on different whole genome amplification methods followed by NGS on the Ion GeneStudio™ S5 Prime System (ThermoFisher Scientific), will be compared. The concordance between cfDNA and trophectoderm biopsies will be evaluated for approximately 150 blastocysts with the best performing niPGT-A protocol.

Selection of the embryo(s) for transfer will be based solely on the PGT-A result from the biopsied cells. The patient's IVF+PGT-A treatment plan and timeline will not be altered.

Sex: Female

Ages: 18 to 46 years old

Volunteers: Accepts Healthy Volunteers

Inclusion criteria

• Patients undergoing fertility treatment with PGT-A (Recombinant FSH antagonist protocol with dual trigger)
• Secondary infertility
• BMI 18- 35 kg/m2
• Sperm: fresh ejaculated sperm (abstinence: 2-3 days)
• At least one blastocyst biopsied on day 6

Exclusion criteria

• High progesterone on day of trigger (>1.5ng/ml)
• Vitrified oocytes
• Frozen sperm
• Indications for PGT-SR and PGT-M

Trial design

0 participants in 1 patient group

Two types of samples (TE and SCM) will be collected for all blastocysts included in the study

Treatment:

Diagnostic Test: PGT-A / niPGT-A