Research article

Hematopoietic chimerism after allogeneic stem cell transplantation: a comparison of quantitative analysis by automated DNA sizing and fluorescent in situ hybridization

Justyna Jółkowska¹ , Anna Pieczonka² , Tomasz Strabel³ , Dariusz Boruczkowski² , Jacek Wachowiak² , Peter Bader⁴ and Michał Witt^1,5

¹Division of Molecular and Clinical Genetics, Institute of Human Genetics, Strzeszyñska 32, 60-479 Poznañ, Poland

²Department of Pediatric Oncology, Hematology and Transplantology, University of Medical Sciences, Szpitalna 27/33, 60-572 Poznañ, Poland

³University of Agriculture, Department of Genetics and Animal Breeding, Wołyñska 33, 60-627 Poznañ, Poland

⁴University Children Hospital III MRD-/ Chimerism Laboratory, Frankfurt/Main, Germany (University Children Hospital Tuebingen previously)

⁵International Institute of Molecular and Cell Biology, Trojdena 4, Warszawa, Poland

author email corresponding author email

BMC Blood Disorders 2005, 5:1doi:10.1186/1471-2326-5-1

Published:	10 January 2005

Abstract

Background

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias. In the context of the significant risk of graft failure after allo-HSCT from alternative donors and the risk of relapse in recipients transplanted for malignancy, the precise monitoring of posttransplant hematopoietic chimerism is of utmost interest. Useful molecular methods for chimerism quantification after allogeneic transplantation, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers. Such analyses can be performed regardless of donor's and recipient's sex. Additionally, in patients after sex-mismatched allo-HSCT, fluorescent in situ hybridization (FISH) can be applied.

Methods

We compared different techniques for analysis of posttransplant chimerism, namely FISH and PCR-based molecular methods with automated detection of fluorescent products in an ALFExpress DNA Sequencer (Pharmacia) or ABI 310 Genetic Analyzer (PE). We used Spearman correlation test.

Results

We have found high correlation between results obtained from the PCR/ALF Express and PCR/ABI 310 Genetic Analyzer. Lower, but still positive correlations were found between results of FISH technique and results obtained using automated DNA sizing technology.

Conclusions

All the methods applied enable a rapid and accurate detection of post-HSCT chimerism.