Demonstration of chimerism after allogeneic bone marrow transplantation by polymerase chain reaction of Y-chromosome-specific nucleotide sequences--characterization of a new technical approach.
Various techniques are applied to assess chimerism after allogeneic bone marrow transplantation. The polymerase chain reaction (PCR) of donor- and/or host-specific gene sequences provides a rapid and highly sensitive technique. We describe the characterization and application of PCR for the amplification of Y-chromosome-specific DNA in blood cells recovered from stored slides. Four different primer pair combinations were used. PCR can be rapidly performed on stained or unstained slide material with varying sensitivity--depending on the primer combination. The lowest limit of detection is one male cell in 500-1000 female cells. The technique was applied to follow the early post-transplant course of 15 male patients who received grafts from female donors and found a high incidence of mixed chimerism during the first three months after BMT and a striking fluctuation between positive and negative results in the follow-up of individual patients. We conclude that PCR for the detection of male-specific DNA sequences can be successfully performed with high sensitivity on material recovered from stored blood slides.
Wilborn
F
F
Abt. Hämatologie, Klinikum Rudolf Virchow, Freie Universität Berlin, Germany.
Schmidt
C A
CA
Siegert
W
W
D000293:Adolescent; D000328:Adult; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000473:pathology*; D005260:Female; D006801:Humans; D007938:Leukemia / Q000601:surgery; D008297:Male; D008875:Middle Aged; D009190:Myelodysplastic Syndromes / Q000601:surgery; D016133:Polymerase Chain Reaction; D011828:Radiation Chimera; D013997:Time Factors; D014998:Y Chromosome