Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation

Area of Science:

• Molecular Biology
• Clinical Diagnostics
• Immunology

Background:

• Hematopoietic chimerism monitoring is vital post-allogeneic stem cell transplantation.
• Short tandem repeat (STR) amplification is the current standard but lacks sensitivity (≥1%).
• Digital PCR (dPCR) offers high sensitivity (≤0.1%) and precise quantification.

Purpose of the Study:

• To develop and validate a comprehensive panel of digital PCR assays for routine clinical chimerism assessment.
• To establish a sensitive and accurate alternative to current STR-based methods.

Main Methods:

• Tested 52 deletion/insertion polymorphisms (DIPs) for duplex dPCR assays.
• Optimized and validated 29 high-performance DIPs for chimerism analysis.
• Validated assays on retrospective patient samples alongside STR analysis.

Main Results:

• Identified and validated 29 DIPs suitable for duplex dPCR.
• Established assays demonstrated excellent correlation with STR amplification results.
• Developed a screening plate for convenient assay selection.

Conclusions:

• A comprehensive dPCR system for high-sensitivity hematopoietic chimerism measurement has been established.
• This dPCR system offers a precise and sensitive tool for clinical routine diagnostics.
• The developed assays are a promising alternative to STR analysis for post-transplant monitoring.