Analysis of Human Genomic DNA in Embryo's Culture Media

February 8, 2021 updated by: Palini Simone, Cervesi Hospital, Cattolica, Italy

Analysis of Human Genomic DNA in Embryo's Culture Media Targeting on a Single Gene Disease and Point Mutations

try to find genomic DNA in culture medium after the embryos develop on Day 3 and Day 5 also in single step culture media. using direct PCR Polymerase Chain Reaction and also WGA Whole Genome Amplification before PCR and sequencing of the samples to find the point mutation

Study Overview

Status

Recruiting

Conditions

Intervention / Treatment

Detailed Description

Human fertilized oocytes were individually cultured from Day 1 to Day 3, and from day 3 to days 5/6, in 10 ul medium. A total of 100 samples (50 spent media from Day 3 and 50 from Day 5/6) will be use in the study.

Double-stranded DNA (dsDNA) amplifiability will be evaluated in embryos culture media by quantitative PCR on a multicopy gene target (TBC1D3). Quantification will be made with a standard curve constructed with genomic DNA (with culture media -day 3 and day5-, to take into account media inhibitory effects) TSPY1 amplification will be use to assess the presence of Y chromosome WGA followed by PCR on a single copy gene (MTHFR gene) will be performed on a subset of some samples to monitor the C677T polymorphism (genotyping by sequencing) WGA followed by PCR on a single copy gene (MTHFR) will be also performed on blastocoele fluids

Study Type

Observational

Enrollment (Anticipated)

100

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Locations

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

18 years to 45 years (Adult)

Accepts Healthy Volunteers

No

Genders Eligible for Study

All

Sampling Method

Non-Probability Sample

Study Population

all the mediun descarted after embryos culture in vitro

Description

Inclusion Criteria

  • all the patients undergoing IVF treatment at Cervesi Hospital

Exclusion Criteria:

  • patients with embryos that can't be cultured from day 0 (insemination day) to day 5/6 (blastocyst stage)

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Observational Models: Cohort
  • Time Perspectives: Retrospective

Cohorts and Interventions

Group / Cohort
Intervention / Treatment
In Vitro Fertilization treatment
culture medium discarded after IVF treatment
Genetic: Polymerase Chain Reaction and Whole Genome Amplification
Polymerase Chain Reaction and Whole Genome Amplification

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Time Frame
genomic DNA in culture medium (pg)
Time Frame: 1 year
1 year

Secondary Outcome Measures

Outcome Measure
Measure Description
Time Frame
amplification of point mutation
Time Frame: 1 year
amplification of point mutation with PCR (polymerase chain reaction)
1 year

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Cervesi Hospital, Cattolica, Italy

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

January 12, 2020

Primary Completion (Anticipated)

November 1, 2021

Study Completion (Anticipated)

December 1, 2022

Study Registration Dates

First Submitted

February 3, 2015

First Submitted That Met QC Criteria

February 9, 2015

First Posted (Estimate)

February 10, 2015

Study Record Updates

Last Update Posted (Actual)

February 10, 2021

Last Update Submitted That Met QC Criteria

February 8, 2021

Last Verified

February 1, 2021

More Information

Terms related to this study

Keywords

Other Study ID Numbers

  • PGD on culture media is easy

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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