Detection and quantitation of genetically marked acute myeloid leukemia bycompetitive polymerase chain reaction after autologous bone marrow transplantation: A preclinical model for minimal residual disease

Preclinical models and methods aimed at detecting and quantitating minimal residual disease (MRD) after autologous bone marrow transplantation (BMT) f or acute myeloid leukemia (AML) could facilitate assessment of innovative t herapeutic strategies for their antileukemic potential. Among the various t echniques exploited to identify MRD, polymerase chain reaction (PCR) proved to be a valuable tool in instances in which clonogeneic markers are involv ed during the evolution of disease, In human AML, however, detection of MRD by PCR is limited to a minority of subgroups, as clonospecific markers are absent or presently unknown, Although gene labeling has proved to be effic ient in detecting marker-devoid leukemia cells in preclinical models, detec tion and quantitation by PCR have not yet been considered. We therefore dev eloped an experimental model in which detection and quantitation of genetic ally marked murine AML cells are based on a highly sensitive two-step neste d PCR and competitive PCR protocol, respectively, We further demonstrated i ts applicability to a murine syngeneic BMT model that was designed to monit or minimal numbers of gene-tagged AML cells at various time intervals after transplantation. Our results showed that detection and quantitation could reproducibly be achieved at levels as low as one in 10(6) and 10(5) cells, respectively, (C) 1999 International Society for Experimental Hematology Pu blished by Elsevier Science Inc.