Whole-Genome Amplification on Single Circulating Trophoblast Cell

Area of Science:

• Genetics
• Molecular Biology
• Reproductive Medicine

Background:

• Whole-genome amplification (WGA) is crucial for analyzing limited DNA samples, such as single cells.
• Cell-based noninvasive prenatal testing (cb-NIPT) relies on analyzing fetal DNA from maternal blood.
• Existing WGA methods may conflict with single trophoblast isolation protocols.

Purpose of the Study:

• To provide a detailed protocol for WGA on single circulating trophoblast cells.
• To optimize WGA for compatibility with cb-NIPT workflows.
• To facilitate comprehensive genetic analysis from trace amounts of DNA.

Main Methods:

• Isolation of single circulating trophoblast cells from maternal blood.
• Application of a WGA technique compatible with trophoblast isolation steps (fixation, permeabilization, immunostaining).
• SNP genotyping and next-generation sequencing of amplified DNA.

Main Results:

• Successful amplification of the entire genome from isolated single trophoblast cells.
• Generation of sufficient DNA for downstream genetic analysis.
• Demonstration of a viable protocol for cb-NIPT.

Conclusions:

• This protocol enables WGA on single circulating trophoblasts for cb-NIPT.
• The method supports comprehensive genetic testing from trace DNA.
• It addresses compatibility challenges between WGA and cell isolation techniques.