Molecular assessment of post-BMT chimerism using various biologic specimens and automated DNA sizing technology

Highly polymorphic microsatellite markers provide useful genetic markers fo r detection of complete or mixed chimerism in patients after allogeneic BMT (allo-BMT). We report application of automated DNA sizing technology for d etection of post-BMT chimerism using fresh peripheral blood, BM, or archiva l blood smears and various DNA isolation techniques. Donors' and recipients ' DNA was amplified with fluorescent PCR primers specific for short tandem repeat (STR) marker loci: FGA, VWA, TH01, F13A1, D21S11. Chimerism was asse ssed in 14 recipients after allo-BMT. A complete chimerism was detected in 10 patients, in 3 patients we observed fluctuations of chimerism status, an d mixed chimerism was assessed in 1 patient. We show that DNA from differen t types of biologic specimens (whole peripheral blood, BM suspension, archi val blood smears), prepared according to the various isolation techniques ( salting-out method, phenol chloroform extraction, Chelex procedure) and amp lified with fluorescent PGR primers for microsatellite markers, enable iden tification of chimerism status following allo-BMT in children.