title: Detection of mixed chimaerism in flow-sorted cell subpopulations by PCR-amplified VNTR markers after allogeneic bone marrow transplantation.
abstract: The amplification of Variable Number of Tandem Repeats (VNTR) by the polymerase chain reaction (PCR) was used to determine the extent of chimaerism in flow sorted lymphoid and myeloid cell populations following allogeneic bone marrow transplantation (BMT). Pre-BMT screening with a set of five VNTR revealed that at least one marker was maximally informative in 95% of donor-recipient pairs. Mixing reconstruction experiments indicated that detection of 1-5% of the minor cell population in a sample of 5 x 10(3) nucleated cells is feasible. Flow sorted post-transplant peripheral blood B- and T-lymphocyte, natural killer and monocyte cell populations were subjected to PCR-VNTR marker analysis. It was shown that this procedure can be used for the early detection of engraftment and the identification of mixed chimaerism in various haematopoietic cell lineages in patients with leukaemia or severe combined immune deficiency, treated with allogeneic BMT.
authors.affiliation: MGC-Human Genetics Department, Leiden University, The Netherlands.
authors.forename: J E
authors.initials: JE
authors.lastname: van Leeuwen
authors.forename: M J
authors.initials: MJ
authors.lastname: van Tol
authors.forename: B G
authors.initials: BG
authors.lastname: Bodzinga
authors.forename: J T
authors.initials: JT
authors.lastname: Wijnen
authors.forename: M
authors.initials: M
authors.lastname: van der Keur
authors.forename: A M
authors.initials: AM
authors.lastname: Joosten
authors.forename: H J
authors.initials: HJ
authors.lastname: Tanke
authors.forename: J M
authors.initials: JM
authors.lastname: Vossen
authors.forename: P M
authors.initials: PM
authors.lastname: Khan
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D016026:Bone Marrow Transplantation / Q000473:pathology*; D002469:Cell Separation; D002648:Child; D002675:Child, Preschool; D002678:Chimera / Q000235:genetics*; D005434:Flow Cytometry; D006801:Humans; D007223:Infant; D016131:Lymphocyte Subsets / Q000473:pathology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D014019:Tissue Donors