title: Hematopoietic chimerism after allogeneic stem cell transplantation: a comparison of quantitative analysis by automated DNA sizing and fluorescent in situ hybridization.
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.
authors.affiliation: Division of Molecular and Clinical Genetics, Institute of Human Genetics, Strzeszyñska 32, 60-479 Poznañ, Poland. justynx@man.poznan.pl.
authors.forename: Justyna
authors.initials: J
authors.lastname: Jólkowska
authors.forename: Anna
authors.initials: A
authors.lastname: Pieczonka
authors.forename: Tomasz
authors.initials: T
authors.lastname: Strabel
authors.forename: Dariusz
authors.initials: D
authors.lastname: Boruczkowski
authors.forename: Jacek
authors.initials: J
authors.lastname: Wachowiak
authors.forename: Peter
authors.initials: P
authors.lastname: Bader
authors.forename: Michal
authors.initials: M
authors.lastname: Witt