Generation of quartet reference materials for preimplantation genetic testing for structural rearrangements

Area of Science:

• Genetics
• Reproductive Medicine
• Bioinformatics

Background:

• Preimplantation genetic testing for structural rearrangements (PGT-SR) is crucial for selecting chromosomally normal embryos.
• A significant need exists for reliable reference materials (RMs) to develop and validate PGT-SR methods.

Purpose of the Study:

• To design and produce a novel, renewable panel of reference materials for PGT-SR.
• To establish a quality assurance framework for PGT-SR assays.

Main Methods:

• Developed a strategy using quartet families of structural rearrangements to create renewable RMs.
• Created a panel of RMs including reciprocal translocations, Robertsonian translocations, inversions, and normal families.
• Characterized RMs using DNA and sorted cells, validated by volunteer laboratories using various sequencing platforms.

Main Results:

• Successfully developed a panel of quartet RMs for PGT-SR from 13 diverse genetic families.
• Achieved 100% accuracy and specificity in RM characterization across multiple labs and sequencing platforms.
• Demonstrated long-read genome sequencing's superiority over short-read for complex structural rearrangements.

Conclusions:

• The developed quartet RMs for PGT-SR are well-characterized, renewable, and accessible, vital for quality assurance.
• The methodology supports proficiency testing and enhances structural rearrangement detection accuracy.
• Adaptable methods promise broader application in preimplantation genetic testing, advancing assisted reproduction.