DNA Amplification in Blastocoel Fluid

NCT04744844 | Terminated

• 1 other identifier: BF 13.1.2021:51
• Study Type: interventional
• Target: 15
• Locations: 1 country
• Sites: 1

Brief Summary

Introduction: Although innovative procedural changes in frozen embryo transfer (FET) cycles have increased the implantation rate of blastocysts transferred significantly, blastocyst selection remains a significant limiting factor in implantation outcomes. To improve implantation rates requires conventional microscopic blastocyst morphology scoring/selection technique to be replaced by an enhanced blastocyst selection technique or for the conventional morphology selection technique to be strengthened by novel supplementary selection techniques. Blastocoel fluid biopsy with DNA amplification is a minimally invasive (mi) technique that may supplement a blastocyst morphology score variables with a genetic variable. Objective: In the present randomized controlled trial (RCT), DNA amplification in blastocoel fluid biopsies (BF-biopsy) will be investigated as a supplementary measure to select blastocysts for transfer in conjunction with blastocyst morphology scores. The objective will be to develop a minimally invasive blastocyst selection technique, which will improve selection and increase clinical implantations, while not increasing costs. Materials and Methods: A single IVF centre double-blind randomised controlled trial, with patients recruited having female age 18 to 35 years from infertile patients presenting for freeze-all-IVF treatment. Enrolled patients (N = 500) with ≥five 2PN zygotes after ICSI will be randomised (1:1) to the two arms of the trial (i.e., test and control arm). In the test arm, 3 blastocysts will undergo blastocoel fluid biopsy (BF-biopsy) and whole-genomic amplification. Single blastocysts with no DNA amplification will be transferred in FETs of the test arm and single top-scoring blastocysts will be transferred in FETs of the control arm. The primary outcome measure of the trial will be clinical implantation (i.e., gestational sac with fetal heartbeat). Results: The clinical implantation outcomes of FETs in which score-selected single blastocyst with no DNA amplification and score-selected single blastocysts were transferred will be compared.

Conditions

Clinical Pregnancy; Embryo Implantation

Keywords

DNA; amplification; blastocyst fluid; blastocyst selection; frozen embryo transfer; minimally invasive

Outcome Measures

Primary Outcomes (1)

• clinical implantation

  Clinical implantation will be defined as a cycle with an ultrasound confirmed normal gestational sac and heartbeat.

  Transvaginal ultrasound examinations will be performed after 5 weeks of gestation

Secondary Outcomes (2)

• pregnancy

  Blood serum pregnancy tests will be performed 9 days after blastocyst transfer

• ongoing pregnancy

  Transvaginal ultrasound examinations will be performed after 12 weeks of gestation

Study Arms (2)

DNA-amplification selection

EXPERIMENTAL

In the experimental arm, all blastocysts will undergo routine morphological assessment, with the 3 top-scoring blastocysts undergoing blastocoel fluid biopsy (BF-biopsy) and whole-genomic amplification. A single blastocyst with no DNA amplification will be selected for transfer in a frozen embryo transfer cycle.

Procedure: blastocoel fluid biopsy

Morfological-score selection

ACTIVE COMPARATOR

In the active comparator arm, all blastocysts will undergo routine morphological assessment. The (single) top-scoring blastocyst will be selected for transfer in a frozen embryo transfer cycle.

Procedure: blastocoel fluid biopsy

Interventions

blastocoel fluid biopsy PROCEDURE

In the present study, blastocoel fluid biopsy (BF-biopsy) and collapse will be performed using a microinjection pipette similar to that used to perform ICSI. The pipette will be pushed gently through the zona pellucida and TE, and up to 90% of the BF will be aspirated.

DNA-amplification selection; Morfological-score selection

Eligibility Criteria

• Age: 18 Years - 35 Years
• Sex: female
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• Patients with female age 18≤35 years on the day of consultation (with the clinician projecting female age to be ≤35 on the day of oocyte retrieval).
• Patients who provide informed consent to participate in the trial and for the use of their anonymized data in research.
• Patients with ≤2 previous IVF treatments.
• Patients predicted to have single blastocyst transfers.

You may not qualify if:

• Patients with female age \>35 years on the day of consultation.
• Female patients with insulin-dependent diabetes or non-insulin-dependent diabetes mellitus and female patients with gastrointestinal, cardiovascular, pulmonary, liver or kidney disease.
• Female patients with any contraindications or allergies to the drugs used in routine freeze-all-IVF.
• Patients undergoing conventional PGT-A (aneuploidy) or PGT-M (monogenic disorders)
• Patients with less than 5 2-PN zygotes on day 1 of embryo development will be excluded from randomization.

Sponsors & Collaborators

• Antalya IVF: lead

Study Sites (1)

Antalya IVF

Antalya, 07080, Turkey (Türkiye)

Location

Related Publications (9)

• Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, Nagy ZP, Ubaldi FM. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014 Jun;29(6):1173-81. doi: 10.1093/humrep/deu033. Epub 2014 Feb 26.

  PMID: 24578475 RESULT

• Iwayama H, Hochi S, Yamashita M. In vitro and in vivo viability of human blastocysts collapsed by laser pulse or osmotic shock prior to vitrification. J Assist Reprod Genet. 2011 Apr;28(4):355-61. doi: 10.1007/s10815-010-9522-4. Epub 2010 Dec 9.

  PMID: 21152966 RESULT

• Magli MC, Albanese C, Crippa A, Tabanelli C, Ferraretti AP, Gianaroli L. Deoxyribonucleic acid detection in blastocoelic fluid: a new predictor of embryo ploidy and viable pregnancy. Fertil Steril. 2019 Jan;111(1):77-85. doi: 10.1016/j.fertnstert.2018.09.016. Epub 2018 Dec 5.

  PMID: 30528055 RESULT

• Ozgur K, Berkkanoglu M, Bulut H, Yoruk GDA, Candurmaz NN, Coetzee K. Single best euploid versus single best unknown-ploidy blastocyst frozen embryo transfers: a randomized controlled trial. J Assist Reprod Genet. 2019 Apr;36(4):629-636. doi: 10.1007/s10815-018-01399-1. Epub 2019 Jan 7.

  PMID: 30617927 RESULT

• Palini S, Galluzzi L, De Stefani S, Bianchi M, Wells D, Magnani M, Bulletti C. Genomic DNA in human blastocoele fluid. Reprod Biomed Online. 2013 Jun;26(6):603-10. doi: 10.1016/j.rbmo.2013.02.012. Epub 2013 Mar 13.

  PMID: 23557766 RESULT

• Veeck LL: Atlas of the Human Oocytes and Early Conceptus. 1991, Vol. 2. Baltimore, Williams & Willkins Co.

  RESULT

• Paulson RJ, Reichman DE, Zaninovic N, Goodman LR, Racowsky C. Time-lapse imaging: clearly useful to both laboratory personnel and patient outcomes versus just because we can doesn't mean we should. Fertil Steril. 2018 Apr;109(4):584-591. doi: 10.1016/j.fertnstert.2018.01.042. No abstract available.

  PMID: 29653705 RESULT

• Gleicher N, Metzger J, Croft G, Kushnir VA, Albertini DF, Barad DH. A single trophectoderm biopsy at blastocyst stage is mathematically unable to determine embryo ploidy accurately enough for clinical use. Reprod Biol Endocrinol. 2017 Apr 27;15(1):33. doi: 10.1186/s12958-017-0251-8.

  PMID: 28449669 RESULT

• Mukaida T, Oka C, Goto T, Takahashi K. Artificial shrinkage of blastocoeles using either a micro-needle or a laser pulse prior to the cooling steps of vitrification improves survival rate and pregnancy outcome of vitrified human blastocysts. Hum Reprod. 2006 Dec;21(12):3246-52. doi: 10.1093/humrep/del285. Epub 2006 Aug 26.

  PMID: 16936299 RESULT

Study Officials

• Kemal Ozgur, MD

  Antalya IVF

  STUDY DIRECTOR

• Kevin Coetzee, PhD

  Antalya IVF

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: DOUBLE
• Who Masked: PARTICIPANT, CARE PROVIDER
• Purpose: TREATMENT
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Scientific officer

Model Details: A single IVF centre double-blind randomised controlled trial.

Study Record Dates

• First Submitted: February 4, 2021
• First Posted: February 9, 2021
• Study Start: January 1, 2022
• Primary Completion: January 31, 2024
• Study Completion: January 31, 2024
• Last Updated: February 20, 2024

Record last verified: 2024-02

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, CSR
• Time Frame: To be published immediately after publication acceptance.
• Access Criteria: Open access at the Mendeley Data at the URLs provided

Locations

TU (1)