Analysis of Human Genomic DNA in Embryo's Culture Media

Single-Gene Disease Clinical Trial

Official title:

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

Clinical Trial Details — Status: Recruiting

Administrative data

• NCT number: NCT02359747
• Other study ID #: PGD on culture media is easy
• Status: Recruiting
• Start date: January 12, 2020
• Est. completion date: December 2022

Study information

• Verified date: February 2021
• Source: Cervesi Hospital, Cattolica, Italy
• Contact: Simone Palini
• Phone: +393394572101
• Email: simonepalini[@]yahoo.it
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

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

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

Recruitment information / eligibility

• Status: Recruiting
• Enrollment: 100
• Est. completion date: December 2022
• Est. primary completion date: November 2021
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years to 45 Years

Eligibility

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)

Related Conditions & MeSH terms

• Single-Gene Disease

Intervention

Genetic:
• Polymerase Chain Reaction and Whole Genome Amplification
Polymerase Chain Reaction and Whole Genome Amplification

Locations

Country	Name	City	State
Italy	Cervesi Hospital	Cattolica
Italy	cervesi Hospital	Cattolica

Sponsors (1)

Lead Sponsor	Collaborator
Cervesi Hospital, Cattolica, Italy

Country where clinical trial is conducted

Italy, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	genomic DNA in culture medium (pg)		1 year
Secondary	amplification of point mutation	amplification of point mutation with PCR (polymerase chain reaction)	1 year