Analysis of Human Genomic DNA in Embryo's Culture Media

NCT ID: NCT02359747

Last Updated: 2021-02-10

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

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Recruitment Status

UNKNOWN

Total Enrollment

100 participants

Study Classification

OBSERVATIONAL

Study Start Date

2020-01-12

Study Completion Date

2022-12-31

Brief Summary

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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

Detailed Description

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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

Conditions

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Single-Gene Disease

Study Design

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Observational Model Type

COHORT

Study Time Perspective

RETROSPECTIVE

Study Groups

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In Vitro Fertilization treatment

culture medium discarded after IVF treatment

Polymerase Chain Reaction and Whole Genome Amplification

Intervention Type GENETIC

Polymerase Chain Reaction and Whole Genome Amplification

Interventions

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Polymerase Chain Reaction and Whole Genome Amplification

Polymerase Chain Reaction and Whole Genome Amplification

Intervention Type GENETIC

Eligibility Criteria

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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)
Minimum Eligible Age

18 Years

Maximum Eligible Age

45 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

No

Sponsors

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Cervesi Hospital, Cattolica, Italy

OTHER

Sponsor Role lead

Responsible Party

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Palini Simone

Senior Clinica Embryologist Lab Director

Responsibility Role PRINCIPAL_INVESTIGATOR

Locations

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Cervesi Hospital

Cattolica, , Italy

Site Status RECRUITING

cervesi Hospital

Cattolica, , Italy

Site Status RECRUITING

Countries

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Italy

Central Contacts

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Simone Palini

Role: CONTACT

+393394572101

Facility Contacts

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Luca Galluzzi

Role: primary

silvia de stefani

Role: primary

Other Identifiers

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PGD on culture media is easy

Identifier Type: -

Identifier Source: org_study_id

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