Quantitative analysis of chimerism using a short tandem repeat method on a fluorescent automated DNA sequencer
A L Lobashevsky1, R W Senkbeil, J E Townsend
1Immunology Histocompatibility Laboratory, James Whitcomb Riley Hospital for Children, Indianapolis, IN, 46033, USA. alobashe@iupui.edu
Clinical and Laboratory Haematology
|January 25, 2006
Summary
This study developed a sensitive method for monitoring stem cell transplantation (SCT) engraftment using automated DNA sequencing. The technique accurately detects microchimerism (MC), crucial for managing transplant complications.
Area of Science:
- Hematology
- Molecular Biology
- Genetics
Background:
- Stem cell transplantation (SCT) is a vital treatment for various diseases.
- Monitoring engraftment and microchimerism (MC) is essential for diagnosing relapse, rejection, and graft-versus-host disease (GVHD).
Purpose of the Study:
- To develop a sensitive and straightforward method for MC lineage analysis.
- To utilize the Visible Genetics fluorescence automated sequencer for MC monitoring.
Main Methods:
- Developed a method using polymerase chain reaction (PCR) amplification of short tandem repeats (STRs).
- Analyzed donor/recipient DNA mixtures and specific cell subsets (CD3+, CD19+, CD15+).
- Employed Visible Genetics software for semi-quantitative analysis and calculating donor-specific signal percentage.
Main Results:
- Achieved a sensitivity of 0.8% to 6.2% for DNA and cellular MC detection.
- Demonstrated a strong linear correlation (r = 0.94) between donor cell numbers and MC signal intensity.
- Validated the Visible Genetics sequencer for effective MC analysis.
Conclusions:
- The Visible Genetics sequencer provides a sensitive and reliable tool for MC analysis in SCT recipients.
- This method aids in the early diagnosis and management of SCT complications.
- Facilitates improved patient outcomes through precise engraftment monitoring.
