Analysis of the Composition of the Vaginal Microbiota During IVF Treatment and Correlation With Serum Progesterone Level on the Day of Embryo Transfer

Study Overview
=================
Brief Summary
-----------------
The composition of the vaginal microbiota varies throughout a woman's life and is sensitive to hormonal and environmental factors. Specifically, hormonal treatments necessary in the medically assisted procreation (MAP) processes can influence the vaginal microbiota. New sequencing techniques have been used to characterize the vaginal microbiota, demonstrating that the microbiota could be divided into 5 classes. The composition of the vaginal microbiota seems to have an implication in the evolution of a pregnancy after IVF. It is therefore essential to have more data on the evolution of the vaginal microbiota at the different stages of IVF treatment and to analyze whether this evolution can be predictive of the success of embryo implantation. Good endometrial progesterone impregnation is an essential prerequisite for ensuring embryo implantation. Indeed, supporting the luteal phase through vaginal progesterone is an essential step in IVF protocols to ensure synchronization between endometrial maturation and embryonic age. However, the serum progesterone level on the day of embryo transfer varies widely between patients. Several factors such as age, vaginal mucosa trophicity, estrogen impregnation and sexual activity are known to affect the vaginal absorption of progesterone. The treatments and vaginal examinations performed during the IVF procedure could also have an impact on the constitution of the vaginal microbiota. The hypothesis of this research is that there is an alteration in the vaginal microbiota during the IVF process which may alter the absorption of vaginal progesterone, with an impact on the failure or success of embryo implantation.

Official Title
-----------------
Analyse de la Composition du Microbiote Vaginal à différents Temps de la Prise en Charge en FIV et corrélation Avec le Taux de progestérone sérique le Jour du Transfert d'Embryon

Conditions
-----------------
IVF

Intervention / Treatment
-----------------
* Other: Vaginal swab
* Other: Blood test


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Patient in round 1 or 2 of IVF or IVF-ICSI with transfer of a fresh embryo with a normal preimplantation assessment, without antibiotic treatment in the 3 months preceding the sample. The patient must have given their free and informed consent and signed the consent form The patient must be a member or beneficiary of a health insurance plan Exclusion Criteria: Use of antibiotics in the 3 months preceding the sample. Patient presenting an anomaly in the implantation assessment. Presence of uterine malformation, hydrosalpinx, chronic vaginosis. Patient undergoing a protocol to study endometrial receptivity. Patient of African American, African or Latin American origin (patients born to two parents of Hispanic origin) as people of African and Hispanic origin have a type IV flora vs people of Caucasian origins who have type I, II, III and V of the Ravel classification . Cancellation of the procedure for various reasons (stimulation failure, collection failure, fertilization failure or embryonic segmentation). Patient taking a freeze-all approach (freezing of the embryos) for various causes (risk of ovarian hyperstimulation, increase in progesterone, too thin endometrium, etc.). The subject is participating in a category 1 interventional study, or is in a period of exclusion determined by a previous study The subject refuses to sign the consent It is impossible to give the subject informed information The patient is under safeguard of justice or state guardianship

Ages Eligible for Study
-----------------
Minimum Age: 18 Years
Maximum Age: 40 Years

Sexes Eligible for Study
-----------------
Female

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

Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Women undergoing IVF<br> | Other: Vaginal swab<br>* Vaginal flora swab 3 months before stimulation, at oocyte puncture, embryo transfer and beta-human chorionic gonadotropin dosing<br>Other: Blood test<br>* Blood sample to measure plasma dose of progesterone<br>|

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

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Class of vaginal flora according to community state type | Classified into 5 classes of vaginal flora described according to Smith & Ravel 2017 | 3 Months before ovarian stimulation | 
| Microbiota richness of vaginal flora | Measured by operational taxonomic unit | 3 Months before ovarian stimulation | 
| Microbiota alpha diversity of vaginal flora | Shannon Index | 3 Months before ovarian stimulation | 
| Microbiota beta diversity of vaginal flora | Bray Curtis index | 3 Months before ovarian stimulation | 
| Plasma progesterone level on the day of embryo transfer | Measured by chemiluminescence (nmol/L) | Day of embryo transfer (Day 0) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Class of vaginal flora according to community state type | Classified into 5 classes of vaginal flora described according to Smith & Ravel 2017 | Day of oocyte puncture (Month 3) | 
| Class of vaginal flora according to community state type | Classified into 5 classes of vaginal flora described according to Smith & Ravel 2017 | Day of embryo transfer (Day 93) | 
| Class of vaginal flora according to community state type | Classified into 5 classes of vaginal flora described according to Smith & Ravel 2017 | Day of beta-human chorionic gonadotropin dosage (Day 108) | 
| Microbiota richness of vaginal flora | Measured by operational taxonomic unit | Day of oocyte puncture (Month 3) | 
| Microbiota richness of vaginal flora | Measured by operational taxonomic unit | Day of embryo transfer (Day 93) | 
| Microbiota richness of vaginal flora | Measured by operational taxonomic unit | Day of beta-human chorionic gonadotropin dosage (Day 108) | 
| Microbiota alpha diversity of vaginal flora | Shannon Index | Day of oocyte puncture (Month 3) | 
| Microbiota alpha diversity of vaginal flora | Shannon Index | Day of embryo transfer (Day 93) | 
| Microbiota alpha diversity of vaginal flora | Shannon Index | Day of beta-human chorionic gonadotropin dosage (Day 108) | 
| Microbiota beta diversity of vaginal flora | Bray Curtis index | Day of oocyte puncture (Month 3) | 
| Microbiota beta diversity of vaginal flora | Bray Curtis index | Day of embryo transfer (Day 93) | 
| Microbiota beta diversity of vaginal flora | Bray Curtis index | Day of beta-human chorionic gonadotropin dosage (Day 108) | 
| Pregnancy success | Yes/No | Month 3 | 
| Pregnancy progression | Implantation failure/progression/miscarriage | Month 3 | 
| Treatments received for IVF | Description of treatments, taken from patient file | Month 3 | 
| Plasma progesterone level | Measured by chemiluminescence (nmol/L) | Month 3 | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
Pregnancy, vaginal microbiota, Progesterone