Analysis of Human Genomic DNA in Embryo's Culture Media

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

Status

UNKNOWN

Phases

Unknown

Study type

Observational

Source

ClinicalTrials.gov

Registry ID

NCT02359747

Enrollment

100

Registered

2015-02-10

Start date

2020-01-12

Completion date

2022-12-31

Last updated

2021-02-10

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

Single-Gene Disease

Keywords

free gDNA, culture media

Brief 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

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

Interventions

GENETICPolymerase Chain Reaction and Whole Genome Amplification

Polymerase Chain Reaction and Whole Genome Amplification

Study design

Observational model

COHORT

Time perspective

RETROSPECTIVE

Eligibility

Sex/Gender

ALL

Age

18 Years to 45 Years

Healthy volunteers

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)

Design outcomes

Primary

Measure	Time frame
genomic DNA in culture medium (pg)	1 year

Secondary

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

Countries

Italy

Contacts

Primary ContactSimone Palini

simonepalini@yahoo.it+393394572101

Outcome results

None listed

Source: ClinicalTrials.gov · Data processed: Feb 4, 2026