Noninvasive Preimplantation Genetic Testing for Aneuploidy

Study Overview
=================
Brief Summary
-----------------
The study is conducted to evaluate the sensitivity and specificity of non-invasive preimplantation genetic testing for aneuploidy (NiPGT-A) using cell free DNA into spent culture medium (SCM).

Detailed Description
-----------------
Our study is a diagnostic study, recruit 218 biopsied blastocysts from couples undergoing intracytoplasmic sperm injection (ICSI) with Preimplantation genetic testing for aneuploidy (PGT-A) or Preimplantation genetic testing for structural rearrangements (PGT-SR) indication. Oocyte retrieval, ICSI and embryo culture are performed by routine protocol. On day 3, each embryo was washed and cultured in single droplets. TE biopsy was conducted and 10μl SCM of biopsied embryos are collected on day 5 or day 6. Both sample types undergo whole-genome amplification (WGA) using Quanti Flour ONE dsDNA system and DNA analysis by next-generation sequencing (NGS) using NextSeq High output 150 cycles kit. Comparison of NGS results from trophectoderm (TE) biopsy and SCM is done to identify the sensitivity and specificity of the test. A sample size of 218 day 5/day 6 embryos will be needed.

Official Title
-----------------
Noninvasive Preimplantation Genetic Testing for Aneuploidy Using Cell Free DNA on Spent Culture Media

Conditions
-----------------
Preimplantation Genetic Testing (PGT), ICSI

Intervention / Treatment
-----------------
* Diagnostic Test: NiPGT-A


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Assisted Reproductive Treatment with ICSI indication Having PGT-A or PGT-SR indication Having an agreement to be enrolled into NiPGT-A study Having blastocyst to biopsy Exclusion Criteria: In vitro maturation cycle
Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Patients with PGT indication<br>Patients undergo in vitro fertilization with PGT-A or for PGT-SR indication | Diagnostic Test: NiPGT-A<br>* NiPGT-A is the technique to screen the aneuploidy status of an embryo without invasion technique, such as biopsy. We use SCM to isolate cfDNA, then amplify and analyse the aneuploidy of embryo secreting this material using NGS technique.<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sensitivity of NiPGT-A | The ability to detect euploid embryo | 4 months after recruiting | 
| Specificity of NiPGT-A | The ability to detect aneuploid embryo | 4 months after recruiting | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Successful amplification rate | Rate of SCM samples with DNA concentration after WGA greater than or equal 1ng/μl | 7 days after SCM collection | 
| Total concordance | The match NGS results from TE biopsy and SCM for ploidy and sex | 14 days after SCM collection | 
| Positive pregnancy rate | Rate of participants with serum human chorionic gonadotropin level greater than 25 mIU/mL | at 2 weeks after embryo transfer | 
| Clinical pregnancy rate | Rate of participants with at least one gestational sac on ultrasound at 7 weeks' gestation with the detection of heart beat activity. | 5 weeks after embryo transfer | 
| Ongoing pregnancy rate | Rate of participants with detectable heart at 12 weeks' gestation or beyond, after the completion of the first transfer | At 12 weeks' gestation | 
| Implantation rate | as the number of gestational sacs per number of embryos transferred | 3 weeks after embryo transfer | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
Cell free DNA (cfDNA), SCM, Sensitivity, NiPGT-A