The development of an indel panel for microchimerism detection.

Area of Science:

• Reproductive immunology
• Molecular diagnostics
• Genetics

Background:

• Microchimerism, the exchange of cells between mother and fetus, plays a role in various physiological and pathological conditions.
• Existing methods for microchimerism detection often rely on sex-mismatch or HLA genotyping, limiting their applicability.

Purpose of the Study:

• To develop a simple, sex-independent assay for microchimerism detection without the need for HLA genotyping.
• To evaluate the efficacy of a novel insertion/deletion (indel) based assay compared to a Y-chromosome marker assay.

Main Methods:

• A multiplex PCR assay detecting insertion/deletion (indel) polymorphisms was developed.
• Fragment analysis by capillary electrophoresis and probe-based qPCR were employed for detection.
• 192 samples from 97 female patients with recurrent pregnancy loss were analyzed using the indel assay and a DYS14 (Y-chromosome) assay.

Main Results:

• The indel method detected microchimerism in 32% of samples, while the DYS14 assay detected it in 29%.
• An overall agreement of 64% was observed between the two methods.
• No statistically significant differences in microchimerism prevalence were found between the methods at different sampling times.

Conclusions:

• The indel assay efficiently detects fetal microchimerism in maternal samples.
• This method allows for the detection of both female and male microchimerism without sex bias or HLA genotyping.
• The indel assay has the potential for routine implementation in clinical diagnostics and research.